LOCUS       sequence-257186-        6346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tetracycline resistance reporter of ChnR activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6346)
  AUTHORS   Thompson MG, Sedaghatian N, Barajas JF, Wehrs M, Bailey CB, Kaplan 
            N, Hillson NJ, Mukhopadhyay A, Keasling JD
  TITLE     Isolation and characterization of novel mutations in the pSC101 
            origin that increase copy number.
  JOURNAL   Sci Rep. 2018 Jan 25;8(1):1590. doi: 10.1038/s41598-018-20016-w.
  PUBMED    29371642
REFERENCE   2  (bases 1 to 6346)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2018-01-25"; volume: "8(1):1590. doi"; pages: " 
            10.1038/s41598-018-20016-w"
COMMENT     Sequence Label: pMGT1
FEATURES             Location/Qualifiers
     source          1..6346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             272..1063
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(1612..2562)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     promoter        complement(4252..4329)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4535..4554
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5147..6346
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
ORIGIN
        1 ccgttgatcg tgctatgatc gactgatgtc atcagcggtg gagtgcaatg tcatgcgctc
       61 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
      121 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
      181 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
      241 gggcagtcgc cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc
      301 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
      361 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
      421 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
      481 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
      541 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
      601 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
      661 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
      721 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
      781 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
      841 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
      901 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
      961 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     1021 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taattgtctt tcttcagctc
     1081 gctgatgata tgcgtacggg ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt
     1141 tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagcg ctatttcttc
     1201 cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc
     1261 tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta
     1321 acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct
     1381 gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca
     1441 gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt
     1501 tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt
     1561 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct
     1621 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     1681 cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt
     1741 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta
     1801 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     1861 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     1921 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     1981 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     2041 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     2101 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     2161 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     2221 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     2281 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     2341 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     2401 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     2461 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     2521 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     2581 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt
     2641 cccgggtgat ctgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     2701 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     2761 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     2821 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     2881 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     2941 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     3001 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     3061 actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg
     3121 tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg
     3181 ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac
     3241 aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc
     3301 gtcttactgt ccctagtcaa tagaggagac tgaattttca gacacgagat tggtgtgttc
     3361 ctgtctgatt tcgatgggtg ttttcccaaa aaatttttga aatactcggg tcatatggct
     3421 ctgatcgcaa aaaccggctt cactggcaat cactttcagt ggaatttctt tttttgaagt
     3481 gagcaaacgt ttagcaaatt gcaccctgag attcatgatg taggcagcag gggaattacc
     3541 aaaatcgact ttaaattttc tcattaaatg aggcacgctc atcttgacta atcccgctaa
     3601 atcttcgagg gtaattttaa tgctcatgtt ttcattgata aattctaata acagtctgcg
     3661 ttgtagggga gtaaaaccag aacggcactg tccttcctta aaatctaatt tggcatactg
     3721 acggagtaaa tgcagggcga tctggttttt aatgctctct aaatacaggt ttccacctaa
     3781 accaccttgc tttaattcta gttctaaaag tgaagtcaga gcaggtaaaa catgatcttc
     3841 cacaccgact tcgtctctga ttcggatcga agcgatatca taatcaaaca ccttattcgc
     3901 cattgaggtg atggagtcat gcgaaatgta gacatgggtg acggcaatat ggtcattcca
     3961 cgcccagcgg gaatcttcac cacaggtcaa caaggagaca taacccggct taaccacttg
     4021 ggtatcccaa gaggcatcgc cttttctacg catttccgcc gcactttttt tatagttgac
     4081 gatcatgtag tcacgcatgg caggaatatg cacatcgagg ttggtatatt cgtaccctct
     4141 gattgcaatg ttttcccaac cggcattcat gctgtcaatt gtgacattac ccggaatcca
     4201 taacacgaca tcttcgggtt tgatcagcgt atttgctttg tctgtgctca tattcaccac
     4261 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     4321 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     4381 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     4441 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     4501 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     4561 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgagt
     4621 ggccgcggga tatcactagt gcggccggtg gtggtggtcc aaactatccg aatatgtaag
     4681 tctaatcact tcaaggaaga ggtttcccat ttcccttctt tctagcagat gaagaagctt
     4741 gcaactaaaa gagattgttt ggatcagtta cccaaaatcg ttgaaaagat tttaactctt
     4801 cgatttttat tttttaggta atcctagccc tctcgggggc taggattaaa aattttaagt
     4861 tattccaaca cgaatgacaa attgttcaat gcaaaataaa aacatacaat atataaatat
     4921 attttttaat taaaacataa gattacaata aaataagaat ttttatttgg agtttgtttt
     4981 ttttctacaa tgatcattat gtacaatttt taggttcacc ccatccaagc cttgtgattg
     5041 cattcctgcg attctttatt caatgaataa gcaatgctat taatcagcaa tgaataacca
     5101 gcactgcaga ttttgaataa attcacatgt cgtaatggag ggtaccatga aacccaacat
     5161 acccctgatc gtaattctga gcactgtcgc gctcgacgct gtcggcatcg gcctgattat
     5221 gccggtgctg ccgggcctcc tgcgcgatct ggttcactcg aacgacgtca ccgcccacta
     5281 tggcattctg ctggcgctgt atgcgttggt gcaatttgcc tgcgcacctg tgctgggcgc
     5341 gctgtcggat cgtttcgggc ggcggccaat cttgctcgtc tcgctggccg gcgccactgt
     5401 cgactacgcc atcatggcga cagcgccttt cctttgggtt ctctatatcg ggcggatcgt
     5461 ggccggcatc accggggcga ctggggcggt agccggcgct tatattgccg atatcactga
     5521 tggcgatgag cgcgcgcggc acttcggctt catgagcgcc tgtttcgggt tcgggatggt
     5581 cgcgggacct gtgctcggtg ggctgatggg cggtttctcc ccccacgctc cgttcttcgc
     5641 cgcggcagcc ttgaacggcc tcaatttcct gacgggctgt ttccttttgc cggagtcgca
     5701 caaaggcgaa cgccggccgt tacgccggga ggctctcaac ccgctcgctt cgttccggtg
     5761 ggcccggggc atgaccgtcg tcgccgccct gatggcggtc ttcttcatca tgcaacttgt
     5821 cggacaggtg ccggccgcgc tttgggtcat tttcggcgag gatcgctttc actgggacgc
     5881 gaccacgatc ggcatttcgc ttgccgcatt tggcattctg cattcactcg cccaggcaat
     5941 gatcaccggc cctgtagccg cccggctcgg cgaaaggcgg gcactcatgc tcggaatgat
     6001 tgccgacggc acaggctaca tcctgcttgc cttcgcgaca cggggatgga tggcgttccc
     6061 gatcatggtc ctgcttgctt cgggtggcat cggaatgccg gcgctgcaag caatgttgtc
     6121 caggcaggtg gatgaggaac gtcaggggca gctgcaaggc tcactggcgg cgctcaccag
     6181 cctgacctcg atcgtcggac ccctcctctt cacggcgatc tatgcggctt ctataacaac
     6241 gtggaacggg tgggcatgga ttgcaggcgc tgccctctac ttgctctgcc tgccggcgct
     6301 gcgtcgcggg ctttggagcg gcgcagggca acgagccgat cgctaa
//
