LOCUS       sequence-64397-e        7965 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 7965)
  AUTHORS   Marx CJ, Lidstrom ME
  TITLE     Development of improved versatile broad-host-range vectors for use 
            in methylotrophs and other Gram-negative bacteria.
  JOURNAL   Microbiology. 2001 Aug;147(Pt 8):2065-75.
  PUBMED    11495985
REFERENCE   2  (bases 1 to 7965)
  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: 
            "Microbiology."; date: "2001-08"; volume: "147(Pt 8)"; pages: 
            "2065-75"
COMMENT     Sequence Label: pCM130
FEATURES             Location/Qualifiers
     source          1..7965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1594..1613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2865..2892)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2984..3070)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3223..3240
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     primer_bind     complement(3252..3269)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3252..3268)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3261..3283)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3477..3496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3596..3618
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3656..3674)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             4460..5659
                     /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(5890..7038)
                     /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(7310..7681)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7714..7823
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atggtacccg ggagctccat caggtgagca cggtcatgaa tcgttcgttg
     1801 agaatgcggt cgtggtaaaa gatcgccttg cccagctggt cggtggtcca ggtcaccggt
     1861 ttgtggtccg ggtagttgta atctcccccg cagaacactt cgttgcggtt accggacggg
     1921 tcgaagaagt agatggtctt gccgtgagtg aggccgtggc gggttgggcc gatatcgata
     1981 gatgtgtcgg tcatggagat caggtcggcg gcgcgaagca agtcttccca ggtttcgagg
     2041 tggaaggaca catgatggag gcggcctttt tccggatggt gaatgaaggc cacgtcgtgg
     2101 gccttggtcg acagactgag aaactgggcg acgcgcgtgc cattttcgtc cagcacctgt
     2161 tcggccagat agaaaccgag caccttggtg aacaggtcat aggtcgccgg caattcgtcg
     2221 ccatacatga gggcgtggtc gaaacgcaca gccgccatac ctttcagatc gcgcggccat
     2281 gcctcgggat tgacgtcatt caaaccccac tttccagtat attccttgtc tgcatacaac
     2341 tcgaagtgat gcccggaggg ggcctggaag cgcacgcgcc ggccacaact gttcagttca
     2401 cctgcgggta gctgctcaac ggcacagcca tatgccatca gatcccgctc cagttgccgg
     2461 agagcatcct catccacaac cttgaaaccc ataaaatcca tgcccggctc gtcagcctcg
     2521 cgtagcacca gggaaaactt atccacttcg gtccaagcct tcagatagac acggccctgg
     2581 tcgtcacggt ccatctcgat caggcccagc aactcgacgt agtgttccag ggccttgctc
     2641 atgtccagta cacgcagctg cacatggccc ggtcgcatta cacctttgtt catgacgtca
     2701 cctcctgaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga
     2761 atctagaatt ccctgctttc ctgatgcaaa aacgaggcta gtttaccgta tctgtggggg
     2821 gatggcttgt agatatgacg acaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     2881 ggatggcctt ctgcttaatt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     2941 ctccgggccg ttgcttcgca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3001 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     3061 gttttatttg atgcctggca gttccctact ctcgcatggg gagaccccac actaccatcg
     3121 gcgctacggc gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg
     3181 ccgccaggca aattctgttt tatcagaccg cttctgcgtt ctgatttaat ctgtatcagg
     3241 ctgaaaaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     3301 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3361 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     3421 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     3481 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     3541 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3601 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     3661 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gcaccctttc
     3721 tcggtccttc aacgttcctg acaacgagcc tccttttcgc caatccatcg acaatcaccg
     3781 cgagtccctg ctcgaacgct gcgtccggac cggcttcgtc gaaggcgtct atcgcggccc
     3841 gcaacagcgg cgagagcgga gcctgttcaa cggtgccgcc gcgctcgccg gcatcgctgt
     3901 cgccggcctg ctcctcaagc acggccccaa cagtgaagta gctgattgtc atcagcgcat
     3961 tgacggcgtc cccggccgaa aaacccgcct cgcagaggaa gcgaagctgc gcgtcggccg
     4021 tttccatctg cggtgcgccc ggtcgcgtgc cggcatggat gcgcgcgcca tcgcggtagg
     4081 cgagcagcgc ctgcctgaag ctgcgggcat tcccgatcag aaatgagcgc cagtcgtcgt
     4141 cggctctcgg caccgaatgc gtatgattct ccgccagcat ggcttcggcc agtgcgtcga
     4201 gcagcgcccg cttgttcctg aagtgccagt aaagcgccgg ctgctgaacc cccaaccgtt
     4261 ccgccagttt gcgtgtcgtc agaccgtcta cgccgacctc gttcaacagg tccagggcgg
     4321 cacggatcac tgtattcggc tgcaactttg tcatgattga cactttatca ctgataaaca
     4381 taatatgtcc accaacttat cagtgataaa gaatccgcgc gttcaatcgg accagcggag
     4441 gctggtccgg aggccagacg tgaaacccaa catacccctg atcgtaattc tgagcactgt
     4501 cgcgctcgac gctgtcggca tcggcctgat tatgccggtg ctgccgggcc tcctgcgcga
     4561 tctggttcac tcgaacgacg tcaccgccca ctatggcatt ctgctggcgc tgtatgcgtt
     4621 ggtgcaattt gcctgcgcac ctgtgctggg cgcgctgtcg gatcgtttcg ggcggcggcc
     4681 aatcttgctc gtctcgctgg ccggcgccac tgtcgactac gccatcatgg cgacagcgcc
     4741 tttcctttgg gttctctata tcgggcggat cgtggccggc atcaccgggg cgactggggc
     4801 ggtagccggc gcttatattg ccgatatcac tgatggcgat gagcgcgcgc ggcacttcgg
     4861 cttcatgagc gcctgtttcg ggttcgggat ggtcgcggga cctgtgctcg gtgggctgat
     4921 gggcggtttc tccccccacg ctccgttctt cgccgcggca gccttgaacg gcctcaattt
     4981 cctgacgggc tgtttccttt tgccggagtc gcacaaaggc gaacgccggc cgttacgccg
     5041 ggaggctctc aacccgctcg cttcgttccg gtgggcccgg ggcatgaccg tcgtcgccgc
     5101 cctgatggcg gtcttcttca tcatgcaact tgtcggacag gtgccggccg cgctttgggt
     5161 cattttcggc gaggatcgct ttcactggga cgcgaccacg atcggcattt cgcttgccgc
     5221 atttggcatt ctgcattcac tcgcccaggc aatgatcacc ggccctgtag ccgcccggct
     5281 cggcgaaagg cgggcactca tgctcggaat gattgccgac ggcacaggct acatcctgct
     5341 tgccttcgcg acacggggat ggatggcgtt cccgatcatg gtcctgcttg cttcgggtgg
     5401 catcggaatg ccggcgctgc aagcaatgtt gtccaggcag gtggatgagg aacgtcaggg
     5461 gcagctgcaa ggctcactgg cggcgctcac cagcctgacc tcgatcgtcg gacccctcct
     5521 cttcacggcg atctatgcgg cttctataac aacgtggaac gggtgggcat ggattgcagg
     5581 cgctgccctc tacttgctct gcctgccggc gctgcgtcgc gggctttgga gcggcgcagg
     5641 gcaacgagcc gatcgctgat cgtggaaacg ataggcctat gccatgcggg tcaaggcgac
     5701 ttccggcaag ctatacgcgc cctagaattg tcaattttaa tcctctgttt atcggcagtt
     5761 cgtagagcgc gccgtgcgtc ccgagcgata ctgagcgaag caagtgcgtc gagcagtgcc
     5821 cgcttgttcc tgaaatgcca gtaaagcgct ggctgctgaa cccccagccg gaactgaccc
     5881 cacaaggccc tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag
     5941 gccgctgcct cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg
     6001 ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg
     6061 gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc
     6121 ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag
     6181 gacctggcaa cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga
     6241 caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg
     6301 cgacaggcat tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg
     6361 gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc
     6421 caacacctgc tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt
     6481 gatcttcacg tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat
     6541 tttcttgttg cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat
     6601 cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt
     6661 cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc
     6721 ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc
     6781 caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc acggcggccg atggcgcggg
     6841 cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac
     6901 catcgagccg acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat
     6961 ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg
     7021 gtcaaacgtc cgattcattc accctccttg cgggattgcc ccgactcacg ccggggcaat
     7081 gtgcccttat tcctgatttg acccgcctgg tgccttggtg tccagataat ccaccttatc
     7141 ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc tcgtacttgg tattccgaat
     7201 cttgccctgc acgaatacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac
     7261 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg
     7321 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct
     7381 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
     7441 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
     7501 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
     7561 aggcctacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
     7621 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     7681 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     7741 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     7801 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     7861 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     7921 ctataatgac cccgaagcag ggttatgcag cggaaaagat ccgtc
//
