LOCUS       pmih155        8588 bp DNA     circular SYN 09-FEB-2023
DEFINITION  ADGRE5(EGF1-5)-mCitrine(CTF).
ACCESSION   .
VERSION     .
KEYWORDS    E5(EGF1-5)-mCitr(CTF)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8588)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 9, 2023 from SnapGene Viewer 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    907..966
                     /label=Signal peptide
     misc_feature    979..1095
                     /label=EGF-like 1
     misc_feature    1096..1251
                     /label=EGF-like 2
     misc_feature    1252..1383
                     /label=EGF-like 3
     misc_feature    1384..1530
                     /label=EGF-like 4
     misc_feature    1531..1677
                     /label=EGF-like 5
     misc_feature    2377..2532
                     /label=GPS
     misc_feature    2491..2499
                     /label=Cleavage site
     misc_feature    2563..2631
                     /label=TM1
     misc_feature    2650..2709
                     /label=TM2
     misc_feature    2752..2820
                     /label=TM3
     misc_feature    2857..2925
                     /label=TM4
     misc_feature    2983..3051
                     /label=TM5
     misc_feature    3109..3177
                     /label=TM6
     misc_feature    3205..3273
                     /label=TM7
     CDS             3304..4017
                     /codon_start=1
                     /product="mCitrine (enhanced YFP with monomerizing A206K 
                     mutation (Zacharias et al., 2002))"
                     /label=mCitrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    4114..4125
                     /label=PBM
     promoter        complement(4144..4162)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    4188..4412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4458..4886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4900..5229
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5296..6090
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6264..6385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    6458..6474
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6482..6512)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6527..6548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6836..7421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7592..8452)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8453..8557)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gccaccatgg gaggccgcgt ctttctcgca ttctgtgtct ggctgactct gccgggagct
      961 gaaacccagg actccagggg ctgtgcccgg tggtgccctc agaactcctc gtgtgtcaat
     1021 gccaccgcct gtcgctgcaa tccagggttc agctcttttt ctgagatcat caccaccccg
     1081 acggagactt gtgacgacat caacgagtgt gcaacaccgt cgaaagtgtc atgcggaaaa
     1141 ttctcggact gctggaacac agaggggagc tacgactgcg tgtgcagccc gggatatgag
     1201 cctgtttctg gggcaaaaac attcaagaat gagagcgaga acacctgtca agatgtggac
     1261 gaatgtcagc agaacccaag gctctgtaaa agctacggca cctgcgtcaa cacccttggc
     1321 agctatacct gccagtgcct gcctggcttc aagttcatac ctgaggatcc gaaggtctgc
     1381 acagatgtga atgaatgcac ctccggacaa aacccgtgcc acagctccac ccactgcctc
     1441 aacaacgtgg gcagctatca gtgccgctgc cgcccgggct ggcaaccgat tccggggtcc
     1501 cccaatggcc caaacaatac cgtctgtgaa gatgtggacg agtgcagctc cgggcagcat
     1561 cagtgtgaca gctccaccgt ctgcttcaac accgtgggtt catacagctg ccgctgccgc
     1621 ccaggctgga agcccagaca cggaatcccg aataaccaaa aggacactgt ctgtgaagat
     1681 atgactttct ccacctggac cccgccccct ggagtccaca gccagacgct ttcccgattc
     1741 ttcgacaaag tccaggacct gggcagagac tccaagacaa gctcagccga ggtcaccatc
     1801 cagaatgtca tcaaattggt ggatgaactg atggaagctc ctggagacgt agaggccctg
     1861 gcgccacctg tccggcacct catagccacc cagctgctct caaaccttga agatatcatg
     1921 aggatcctgg ccaagagcct gcctaaaggc cccttcacct acatttcccc ttcgaacaca
     1981 gagctgaccc tgatgatcca ggagcagggg gacaagaacg tcactatggg tcagagcagc
     2041 gcacgcatga agctgaattg ggctgtggca gctggagccg aggatccagg ccccgccgtg
     2101 gcgggcatcc tctccatcca gaacatgacg acattgctgg ccaatgcctc cttgaacctg
     2161 cattccaaga agcaagccga actggaggag atatatgaaa gcagcatccg tggtgtccaa
     2221 ctcagacgcc tctctgccgt caactccatc tttctgagcc acaacaacac caaggaactc
     2281 aactccccca tccttttcgc cttctcccac cttgagtcct ccgatgggga ggcgggaaga
     2341 gaccctcctg ccaaggacgt gatgcctggg ccacggcagg agctgctctg tgccttctgg
     2401 aagagtgaca gcgacagggg agggcactgg gccaccgagg gctgccaggt gctgggcagc
     2461 aagaacggca gcaccacctg ccaatgcagc cacctgagca gctttgcgat ccttatggct
     2521 cattatgacg tggaggactg gaagctgacc ctgatcacca gggtgggact ggcgctgtca
     2581 ctcttctgcc tgctgctgtg catcctcact ttcctgctgg tgcggcccat ccagggctcg
     2641 cgcaccacca tacacctgca cctctgcatc tgcctcttcg tgggctccac catcttcctg
     2701 gccggcatcg agaacgaagg cggccaggtg gggctgcgct gtcgcctggt ggccgggctg
     2761 ctgcactact gtttcctggc cgccttctgc tggatgagcc tcgaaggcct ggagctctac
     2821 tttcttgtgg tgcgcgtgtt ccaaggccag ggcctgagta cgcgctggct ctgcctgatc
     2881 ggctatggcg tgcccctgct catcgtgggc gtctcggctg ccatctacag caagggctac
     2941 ggccgcccca gatactgctg gttggacttt gagcagggct tcctctggag cttcttggga
     3001 cctgtgacct tcatcatttt gtgcaatgct gtcattttcg tgactaccgt ctggaagctc
     3061 actcagaagt tttctgaaat caatccagac atgaagaaat taaagaaggc gagggcgctg
     3121 accatcacgg ccatcgcgca gctcttcctg ttgggctgca cctgggtctt tggcctgttc
     3181 atcttcgacg atcggagctt ggtgctgacc tatgtgttta ccatcctcaa ctgcctgcag
     3241 ggcgccttcc tctacctgct gcactgcctg ctcaacaaga aggttcggga agaataccgg
     3301 aaggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3361 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3421 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3481 ctcgtgacca ccttcggcta cggcctgatg tgcttcgccc gctaccccga ccacatgaag
     3541 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3601 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3661 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3721 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3781 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3841 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3901 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3961 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtgg
     4021 gcctgcctag ttgctggggg gagcaagtac tcagaattca cctccaccac gtctggcact
     4081 ggccacaatc agacccgggc cctcagggca tcagagtccg gcatatgaac tagagggccc
     4141 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     4201 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4261 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4321 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4381 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4441 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4741 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4801 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4861 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4921 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4981 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5041 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5101 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5161 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5221 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5281 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5341 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5401 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5461 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5521 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5581 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5641 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5701 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5761 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5821 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5881 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5941 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6001 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6061 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6121 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6181 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6241 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6301 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6361 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6421 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6481 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6541 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7261 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     7381 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7501 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7561 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7621 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7681 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7741 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7801 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7861 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7921 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7981 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8041 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8101 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8161 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8221 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8281 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8341 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8401 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8461 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8521 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8581 ctgacgtc
//
