LOCUS       pMIH137        8309 bp DNA     circular SYN 07-FEB-2023
DEFINITION  ADGRE5-mCitrine(CTF).
ACCESSION   .
VERSION     .
KEYWORDS    E5-mCitr(CTF)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8309)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 7, 2023 from SnapGene Viewer 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8309
                     /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..1398
                     /label=EGF-like 5
     misc_feature    2098..2253
                     /label=GPS
     misc_feature    2212..2220
                     /label=Cleavage site
     misc_feature    2284..2352
                     /label=TM1
     misc_feature    2371..2430
                     /label=TM2
     misc_feature    2473..2541
                     /label=TM3
     misc_feature    2578..2646
                     /label=TM4
     misc_feature    2704..2772
                     /label=TM5
     misc_feature    2830..2898
                     /label=TM6
     misc_feature    2926..2994
                     /label=TM7
     CDS             3025..3738
                     /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    3835..3846
                     /label=PBM
     promoter        complement(3865..3883)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3909..4133
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4179..4607
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4621..4950
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5017..5811
                     /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    5985..6106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    6179..6195
                     /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(6203..6233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6248..6269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6557..7142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7313..8173)
                     /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(8174..8278)
                     /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 gagtgcagct ccgggcagca tcagtgtgac agctccaccg tctgcttcaa caccgtgggt
     1321 tcatacagct gccgctgccg cccaggctgg aagcccagac acggaatccc gaataaccaa
     1381 aaggacactg tctgtgaaga tatgactttc tccacctgga ccccgccccc tggagtccac
     1441 agccagacgc tttcccgatt cttcgacaaa gtccaggacc tgggcagaga ctccaagaca
     1501 agctcagccg aggtcaccat ccagaatgtc atcaaattgg tggatgaact gatggaagct
     1561 cctggagacg tagaggccct ggcgccacct gtccggcacc tcatagccac ccagctgctc
     1621 tcaaaccttg aagatatcat gaggatcctg gccaagagcc tgcctaaagg ccccttcacc
     1681 tacatttccc cttcgaacac agagctgacc ctgatgatcc aggagcgggg ggacaagaac
     1741 gtcactatgg gtcagagcag cgcacgcatg aagctgaatt gggctgtggc agctggagcc
     1801 gaggatccag gccccgccgt ggcgggcatc ctctccatcc agaacatgac gacattgctg
     1861 gccaatgcct ccttgaacct gcattccaag aagcaagccg aactggagga gatatatgaa
     1921 agcagcatcc gtggtgtcca actcagacgc ctctctgccg tcaactccat ctttctgagc
     1981 cacaacaaca ccaaggaact caactccccc atccttttcg ccttctccca ccttgagtcc
     2041 tccgatgggg aggcgggaag agaccctcct gccaaggacg tgatgcctgg gccacggcag
     2101 gagctgctct gtgccttctg gaagagtgac agcgacaggg gagggcactg ggccaccgag
     2161 ggctgccagg tgctgggcag caagaacggc agcaccacct gccaatgcag ccacctgagc
     2221 agctttgcga tccttatggc tcattatgac gtggaggact ggaagctgac cctgatcacc
     2281 agggtgggac tggcgctgtc actcttctgc ctgctgctgt gcatcctcac tttcctgctg
     2341 gtgcggccca tccagggctc gcgcaccacc atacacctgc acctctgcat ctgcctcttc
     2401 gtgggctcca ccatcttcct ggccggcatc gagaacgaag gcggccaggt ggggctgcgc
     2461 tgtcgcctgg tggccgggct gctgcactac tgtttcctgg ccgccttctg ctggatgagc
     2521 ctcgaaggcc tggagctcta ctttcttgtg gtgcgcgtgt tccaaggcca gggcctgagt
     2581 acgcgctggc tctgcctgat cggctatggc gtgcccctgc tcatcgtggg cgtctcggct
     2641 gccatctaca gcaagggcta cggccgcccc agatactgct ggttggactt tgagcagggc
     2701 ttcctctgga gcttcttggg acctgtgacc ttcatcattt tgtgcaatgc tgtcattttc
     2761 gtgactaccg tctggaagct cactcagaag ttttctgaaa tcaatccaga catgaagaaa
     2821 ttaaagaagg cgagggcgct gaccatcacg gccatcgcgc agctcttcct gttgggctgc
     2881 acctgggtct ttggcctgtt catcttcgac gatcggagct tggtgctgac ctatgtgttt
     2941 accatcctca actgcctgca gggcgccttc ctctacctgc tgcactgcct gctcaacaag
     3001 aaggttcggg aagaataccg gaaggtgagc aagggcgagg agctgttcac cggggtggtg
     3061 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3121 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3181 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgat gtgcttcgcc
     3241 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3301 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3361 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3421 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3481 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3541 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3601 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     3661 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3721 atggacgagc tgtacaagtg ggcctgccta gttgctgggg ggagcaagta ctcagaattc
     3781 acctccacca cgtctggcac tggccacaat cagacccggg ccctcagggc atcagagtcc
     3841 ggcatatgaa ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     3901 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3961 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4021 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4081 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4141 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4201 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4261 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4321 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4381 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4441 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4501 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4561 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4621 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4681 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4741 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4801 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4861 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4921 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4981 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5041 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5101 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5161 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5221 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5281 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5341 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5401 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5461 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5521 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5581 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5641 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5701 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5761 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5821 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5881 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5941 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6001 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6061 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6121 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6181 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6241 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6301 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6361 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6421 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6481 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6541 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6601 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6661 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6721 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6781 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6841 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6901 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6961 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7021 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7081 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7141 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7201 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7261 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7321 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7381 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7441 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7501 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7561 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7621 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7681 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7741 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7801 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7861 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7921 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7981 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8041 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8101 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8161 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8221 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8281 catttccccg aaaagtgcca cctgacgtc
//
