LOCUS       Exported                8043 bp ds-DNA     circular SYN 04-JAN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8043)
  AUTHORS   Alex J H Fedorec
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jan 4, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8043
                     /organism="recombinant plasmid"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     CDS             complement(87..896)
                     /codon_start=1
                     /label=luxR
                     /translation="MIYNTQNLRQTIGKDKEMGMKNINADDTYRIINKIKACRSNNDIN
                     QCLSDMTKMVHCEYYLLAIIYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYS
                     NSNHSPINWNIFENNAVNKKSPNVIKEAKTSGLITGFSFPIHTANNGFGMLSFAHSEKD
                     NYIDSLFLHACMNIPLIVPSLVDNYRKINIANNKSNNDLTKREKECLAWACEGKSSWDI
                     SKILGCSERTVTFHLTNAQMKLNTTNRCQSISKAILTGAIDCPYFKN"
     promoter        948..1065
                     /label=lux promoter
     CDS             1066..1815
                     /codon_start=1
                     /label=sfGFP_ssrA
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKAANDENYALAA"
     CDS             1855..2478
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=tetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             complement(2591..3469)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3496..3780
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             3850..4551
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             4579..5193
                     /codon_start=1
                     /label=luxI_ssrA
                     /note="from pTD103luxI_sfGFP"
                     /translation="MTIMIKKSDFLAIPSEEYKGILSLRYQVFKQRLEWDLVVENNLES
                     DEYDNSNAEYIYACDDTENVSGCWRLLPTTGDYMLKSVFPELLGQQSAPKDPNIVELSR
                     FAVGKNSSKINNSASEITMKLFEAIYKHAVSQGITEYVTVTSTAIERFLKRIKVPCHRI
                     GDKEIHVLGDTKSVVLSMPINEQFKKAVLNAANDENYALAA"
     terminator      5215..5309
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             5449..6264
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            6420..6528
                     /note="Origin of transference region of RP4 plasmid"
     rep_origin      6564..7152
                     /direction=RIGHT
                     /label=pBR322 ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7338..7478
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(7580..7771)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      7951..8037
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt acccgggcac ctcgagttaa tttttaaagt atgggcaatc aattgctcct
      121 gttaaaattg ctttagaaat actttggcag cggtttgttg tattgagttt catttgcgca
      181 ttggttaaat ggaaagtgac agtacgctca ctgcagccta atatttttga aatatcccaa
      241 gagctttttc cttcgcatgc ccacgctaaa cattcttttt ctcttttggt taaatcgttg
      301 tttgatttat tatttgctat atttattttt cgataattat caactagaga aggaacaatt
      361 aatggtatgt tcatacacgc atgtaaaaat aaactatcta tatagttgtc tttttctgaa
      421 tgtgcaaaac taagcattcc gaagccattg ttagccgtat gaatagggaa actaaaccca
      481 gtgataagac ctgatgtttt cgcttcttta attacatttg gagatttttt atttacagca
      541 ttgttttcaa atatattcca attaattggt gaatgattgg agttagaata atctactata
      601 ggatcatatt ttattaaatt agcgtcatca taatattgcc tccatttttt agggtaatta
      661 tctagaattg aaatatcaga tttaaccata gaatgaggat aaatgatcgc gagtaaataa
      721 tattcacaat gtaccatttt agtcatatca gataagcatt gattaatatc attattgctt
      781 ctacaagctt taattttatt aattattctg tatgtgtcgt cggcatttat gtttttcata
      841 cccatctctt tatccttacc tattgtttgt cgcaagtttt gcgtgttata tatcattaaa
      901 acggtaatgg attgacattt gattctaata aattggattt ttgtcacact attgtatcgc
      961 tgggaataca attacttaac ataagcacct gtaggatcgt acaggtttac gcaagaaaat
     1021 ggtttgttat agtcgaatga attcattaaa gaggagaaag gtaccatgag caaaggagaa
     1081 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     1141 aaattttctg tccgtggaga gggtgaaggt gatgctacaa acggaaaact cacccttaaa
     1201 tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac tactctgacc
     1261 tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag
     1321 agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggacc
     1381 tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta
     1441 aagggtattg attttaaaga agatggaaac attcttggac acaaactcga gtacaacttt
     1501 aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc
     1561 aaaattcgcc acaacgttga agatggttcc gttcaactag cagaccatta tcaacaaaat
     1621 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc gacacaatct
     1681 gtcctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact
     1741 gctgctggga ttacacatgg catggatgag ctctacaaag cagcgaacga cgaaaattac
     1801 gcccttgcag cgtaaaagct tagggccccg gcgaagctag ggacgagagc tagcatgtct
     1861 cgtttagata aaagtaaagt gattaacagc gcattagagc tgcttaatga ggtcggaatc
     1921 gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc tacattgtat
     1981 tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat gttagatagg
     2041 caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt acgtaataac
     2101 gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt acatttaggt
     2161 acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt tttatgccaa
     2221 caaggttttt cactagagaa tgcattatat gcactcagcg ctgtggggca ttttacttta
     2281 ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga aacacctact
     2341 actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca ccaaggtgca
     2401 gagccagcct tcttattcgg ccttgaattg atcatctgcg gattagaaaa acaacttaaa
     2461 tgtgaaagtg ggtcttgaga gctcggactg cctgcaggca tgcataatgt gcctgtcaaa
     2521 tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca attgtctgat
     2581 tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
     2641 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
     2701 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
     2761 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
     2821 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
     2881 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
     2941 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
     3001 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
     3061 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
     3121 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
     3181 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
     3241 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
     3301 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
     3361 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
     3421 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
     3481 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
     3541 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     3601 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     3661 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     3721 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     3781 acccgttttt tgggctaaca ggaggaatta accatgggga gccaccatca ccatcaccat
     3841 ggcagatcta tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc
     3901 accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
     3961 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
     4021 ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc
     4081 gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
     4141 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     4201 aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca
     4261 ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga
     4321 aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacgc caagaccaca
     4381 tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac
     4441 agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca
     4501 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatta agaattcatt
     4561 aaagaggaga aaggtaccat gactataatg ataaaaaaat cggatttttt ggcaattcca
     4621 tcggaggagt ataaaggtat tctaagtctt cgttatcaag tgtttaagca aagacttgag
     4681 tgggacttag ttgtagaaaa taaccttgaa tcagatgagt atgataactc aaatgcagaa
     4741 tatatttatg cttgtgatga tactgaaaat gtaagtggat gctggcgttt attacctaca
     4801 acaggtgatt atatgctgaa aagtgttttt cctgaattgc ttggtcaaca gagtgctccc
     4861 aaagatccta atatagtcga attaagtcgt tttgctgtag gtaaaaatag ctcaaagata
     4921 aataactctg ctagtgaaat tacaatgaaa ctatttgaag ctatatataa acacgctgtt
     4981 agtcaaggta ttacagaata tgtaacagta acatcaacag caatagagcg atttttaaag
     5041 cgtattaaag ttccttgtca tcgtattgga gacaaagaaa ttcatgtatt aggtgatact
     5101 aaatcggttg tattgtctat gcctattaat gaacagttta aaaaagcagt cttaaatgca
     5161 gcgaacgacg aaaattacgc ccttgcagcg taaacgcgtg ctagactagt cttggactcc
     5221 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
     5281 gccgccgggc gttttttatt ggtgagaatc caggggtccc caataattac gatttacgta
     5341 tttaaatttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     5401 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     5461 cagcgtgaaa cgagctgtag ccgtccgcgt ctgaacagca acatggatgc ggatctgtat
     5521 ggctataaat gggcgcgtga taacgtgggt cagagcggcg cgaccattta tcgtctgtat
     5581 ggcaaaccgg atgcgccgga actgtttctg aaacatggca aaggcagcgt ggcgaacgat
     5641 gtgaccgatg aaatggtgcg tctgaactgg ctgaccgaat ttatgccgct gccgaccatt
     5701 aaacatttta ttcgcacccc ggatgatgcg tggctgctga ccaccgcgat tccgggcaaa
     5761 accgcgtttc aggtgctgga agaatatccg gatagcggcg aaaacattgt ggatgcgctg
     5821 gccgtgtttc tgcgtcgtct gcatagcatt ccggtgtgca actgcccgtt taacagcgat
     5881 cgtgtgtttc gtctggccca ggcgcagagc cgtatgaaca acggcctggt ggatgcgagc
     5941 gattttgatg atgaacgtaa cggctggccg gtggaacagg tgtggaaaga aatgcataaa
     6001 ctgctgccgt ttagcccgga tagcgtggtg acccacggcg attttagcct ggataacctg
     6061 attttcgatg aaggcaaact gattggctgc attgatgtgg gccgtgtggg cattgcggat
     6121 cgttatcagg atctggccat tctgtggaac tgcctgggcg aatttagccc gagcctgcaa
     6181 aaacgtctgt ttcagaaata tggcattgat aatccggata tgaacaaact gcaatttcat
     6241 ctgatgctgg atgaattttt ctaataatta attggaccgt tgtccttttc cgctgcataa
     6301 ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat
     6361 acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg
     6421 ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat
     6481 tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccgta ggccggcccc
     6541 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6601 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6661 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6721 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6781 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6841 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6901 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6961 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7021 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7081 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7141 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7201 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7261 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7321 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     7381 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7441 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     7501 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     7561 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     7621 tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg
     7681 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     7741 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     7801 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     7861 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     7921 gggggcgcgc ccagctgtct agggcggcgg atttgtccta ctcaggagag cgttcaccga
     7981 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
     8041 cct
//
