LOCUS       sequence-265879-        7909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7909)
  AUTHORS   Marx CJ, Lidstrom ME
  TITLE     Broad-host-range cre-lox system for antibiotic marker recycling in 
            gram-negative bacteria.
  JOURNAL   Biotechniques. 2002 Nov;33(5):1062-7.
  PUBMED    12449384
REFERENCE   2  (bases 1 to 7909)
  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: 
            "Biotechniques."; date: "2002-11"; volume: "33(5)"; pages: "1062-7"
COMMENT     Sequence Label: pCM157
FEATURES             Location/Qualifiers
     source          1..7909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..632
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1095..1683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1584..1603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1837..1854
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1971..1992
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2007..2012,2013..2030,2031..2037)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2045..2061
                     /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)."
     primer_bind     2050..2072
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2069..2085
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2069..2085
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2145..3176
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTTGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(3186..3203)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3186..3202)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3195..3217)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3411..3430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3530..3552
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3590..3608)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             4394..5593
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(5824..6972)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(7244..7615)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7648..7757
                     /note="incP origin of transfer"
ORIGIN
        1 gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg gccctgcaaa
       61 cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc gttgtggata
      121 cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca cttgaggggc
      181 cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc cggcgacgtg
      241 gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat
      301 gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg gcgcgactac
      361 tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca gatgaggggc
      421 gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt gcaagggttt
      481 ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac caatatttat
      541 aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg
      601 tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc ccccaggggc
      661 tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg accacttctg
      721 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
      781 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
      841 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
      901 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
      961 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1021 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1081 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1141 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1201 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1261 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1321 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1381 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1441 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1501 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1561 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1621 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1681 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1741 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     1801 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     1861 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     1921 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     1981 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     2041 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     2101 cttgcatgcc tgcaggtcga ctctagactg caggtaccgt tgacatgtcc aatttactga
     2161 ccgtacacca aaatttgcct gcattaccgg tcgatgcaac gagtgatgag gttcgcaaga
     2221 acctgatgga catgttcagg gatcgccagg cgttttctga gcatacctgg aaaatgcttc
     2281 tgtccgtttg ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa tggtttcccg
     2341 cagaacctga agatgttcgc gattatcttc tatatcttca ggcgcgcggt ctggcagtaa
     2401 aaactatcca gcaacatttg ggccagctaa acatgcttca tcgtcggtcc gggctgccac
     2461 gaccaagtga cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa gaaaacgttg
     2521 atgccggtga acgtgcaaaa caggctctag cgttcgaacg cactgatttc gaccaggttc
     2581 gttcactcat ggaaaatagc gatcgctgcc aggatatacg taatctggca tttctgggga
     2641 ttgcttataa caccctgtta cgtatagccg aaattgccag gatcagggtt aaagatatct
     2701 cacgtactga cggtgggaga atgttaatcc atattggcag aacgaaaacg ctggttagca
     2761 ccacaggtgt agagaaggca cttagcctgg gggtaactaa actggtcgag cgatggattt
     2821 ccgtctctgg tgtagctgat gatccgaata actacctgtt ttgccgggtc agaaaaaatg
     2881 gtgttgccgc gccatctgcc accagccagc tatcaactcg cgccctggaa gggatttttg
     2941 aagcaactca tcgattgatt tacggcgcta aggatgactc tggtcagaga tacctggcct
     3001 ggtctggaca cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct ggagtttcaa
     3061 taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg aactatatcc
     3121 gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc gattagtaag
     3181 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3241 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3301 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     3361 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     3421 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3481 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3541 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3601 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagcaccc tttctcggtc
     3661 cttcaacgtt cctgacaacg agcctccttt tcgccaatcc atcgacaatc accgcgagtc
     3721 cctgctcgaa cgctgcgtcc ggaccggctt cgtcgaaggc gtctatcgcg gcccgcaaca
     3781 gcggcgagag cggagcctgt tcaacggtgc cgccgcgctc gccggcatcg ctgtcgccgg
     3841 cctgctcctc aagcacggcc ccaacagtga agtagctgat tgtcatcagc gcattgacgg
     3901 cgtccccggc cgaaaaaccc gcctcgcaga ggaagcgaag ctgcgcgtcg gccgtttcca
     3961 tctgcggtgc gcccggtcgc gtgccggcat ggatgcgcgc gccatcgcgg taggcgagca
     4021 gcgcctgcct gaagctgcgg gcattcccga tcagaaatga gcgccagtcg tcgtcggctc
     4081 tcggcaccga atgcgtatga ttctccgcca gcatggcttc ggccagtgcg tcgagcagcg
     4141 cccgcttgtt cctgaagtgc cagtaaagcg ccggctgctg aacccccaac cgttccgcca
     4201 gtttgcgtgt cgtcagaccg tctacgccga cctcgttcaa caggtccagg gcggcacgga
     4261 tcactgtatt cggctgcaac tttgtcatgc ttgacacttt atcactgata aacataatat
     4321 gtccaccaac ttatcagtga taaagaatcc gcgcgttcaa tcggaccagc ggaggctggt
     4381 ccggaggcca gacgtgaaac ccaacatacc cctgatcgta attctgagca ctgtcgcgct
     4441 cgacgctgtc ggcatcggcc tgattatgcc ggtgctgccg ggcctcctgc gcgatctggt
     4501 tcactcgaac gacgtcaccg cccactatgg cattctgctg gcgctgtatg cgttggtgca
     4561 atttgcctgc gcacctgtgc tgggcgcgct gtcggatcgt ttcgggcggc ggccaatctt
     4621 gctcgtctcg ctggccggcg ccactgtcga ctacgccatc atggcgacag cgcctttcct
     4681 ttgggttctc tatatcgggc ggatcgtggc cggcatcacc ggggcgactg gggcggtagc
     4741 cggcgcttat attgccgata tcactgatgg cgatgagcgc gcgcggcact tcggcttcat
     4801 gagcgcctgt ttcgggttcg ggatggtcgc gggacctgtg ctcggtgggc tgatgggcgg
     4861 tttctccccc cacgctccgt tcttcgccgc ggcagccttg aacggcctca atttcctgac
     4921 gggctgtttc cttttgccgg agtcgcacaa aggcgaacgc cggccgttac gccgggaggc
     4981 tctcaacccg ctcgcttcgt tccggtgggc ccggggcatg accgtcgtcg ccgccctgat
     5041 ggcggtcttc ttcatcatgc aacttgtcgg acaggtgccg gccgcgcttt gggtcatttt
     5101 cggcgaggat cgctttcact gggacgcgac cacgatcggc atttcgcttg ccgcatttgg
     5161 cattctgcat tcactcgccc aggcaatgat caccggccct gtagccgccc ggctcggcga
     5221 aaggcgggca ctcatgctcg gaatgattgc cgacggcaca ggctacatcc tgcttgcctt
     5281 cgcgacacgg ggatggatgg cgttcccgat catggtcctg cttgcttcgg gtggcatcgg
     5341 aatgccggcg ctgcaagcaa tgttgtccag gcaggtggat gaggaacgtc aggggcagct
     5401 gcaaggctca ctggcggcgc tcaccagcct gacctcgatc gtcggacccc tcctcttcac
     5461 ggcgatctat gcggcttcta taacaacgtg gaacgggtgg gcatggattg caggcgctgc
     5521 cctctacttg ctctgcctgc cggcgctgcg tcgcgggctt tggagcggcg cagggcaacg
     5581 agccgatcgc tgatcgtgga aacgataggc ctatgccatg cgggtcaagg cgacttccgg
     5641 caagctatac gcgccctaga attgtcaatt ttaatcctct gtttatcggc agttcgtaga
     5701 gcgcgccgtg cgtcccgagc gatactgagc gaagcaagtg cgtcgagcag tgcccgcttg
     5761 ttcctgaaat gccagtaaag cgctggctgc tgaaccccca gccggaactg accccacaag
     5821 gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct
     5881 gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga
     5941 atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga
     6001 gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat
     6061 gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg
     6121 gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga
     6181 ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag
     6241 gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag
     6301 ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac
     6361 ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt
     6421 cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt
     6481 gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc
     6541 cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg
     6601 tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt
     6661 ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc
     6721 tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc
     6781 agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga
     6841 gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc
     6901 ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa
     6961 cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc
     7021 ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat
     7081 gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc
     7141 ctgcacgaat accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt
     7201 ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg
     7261 ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca
     7321 gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga
     7381 gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt
     7441 gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggcct
     7501 acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc
     7561 agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct
     7621 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga
     7681 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
     7741 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
     7801 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
     7861 tgaccccgaa gcagggttat gcagcggaaa agatccgtcg accctttcc
//
