LOCUS       Exported                6724 bp DNA     circular SYN 21-JUL-2025
DEFINITION  E. Coli/Gram-positive Bacterial Shuttle Vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6724)
  AUTHORS   LeBlanc DJ, Lee LN, Abu-Al-Jaibat A
  TITLE     Molecular, genetic, and functional analysis of the basic replicon of
            pVA380-1, a plasmid of oral streptococcal origin.
  JOURNAL   Plasmid. 1992 Sep;28(2):130-45.
  PUBMED    1409970
REFERENCE   2  (bases 1 to 6724)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6724)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6724)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6724)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid."; 
            date: "1992-09"; volume: "28(2)"; pages: "130-45"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..83)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1772..2446
                     /codon_start=1
                     /label=Spec
                     /note="spectinomycin adenyltransferase"
                     /translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
                     GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
                     QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
                     MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
                     KL"
     CDS             complement(2702..3667)
                     /codon_start=1
                     /label=Rep
                     /translation="MTGTENQVFKDYSKNGKDRKWRERKLKNIELAGRLERLGYRSFER
                     VYQCAEVLKFIEQQDGTKKLYQSYFCKNKLCPICNWRRSMKYAYQAELVVNEAMKRYPK
                     GRFLFLTLTIKNISGEKLNKSISEIGRAFNRLMKYKKVDKNVIGYLRATEVTYSTEHEN
                     YHPHLHVLLFVKSSYFTGNNTNYISQEEWTKLWAKAMKLDYTPVVDIRTVKAHKRKNLK
                     SAIIETAKYPVKPFDVDTEDVTLFSEMVKERITEDLTNGLHRKRQIGFGKLFKKIKAEL
                     ALDDVEEGNLVQTGAEESAESTGREIVAFWNWDRKNYFVR"
     terminator      5146..5173
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      5305..5348
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(5644..5671)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(5763..5849)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     6323..6339
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6340..6396
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(6409..6425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6433..6449)
                     /label=lac operator
                     /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(6457..6487)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6502..6523)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc cgataaaccc agcgaaccat ttgaggtgat aggtaagatt ataccgaggt
     1261 atgaaaacga gaattggacc tttacagaat tactctatga agcgccatat ttaaaaagct
     1321 accaagacga agaggatgaa gaggatgagg aggcagattg ccttgaatat attgacaata
     1381 ctgataagat aatatatctt ttatatagaa gatttttcgt tcgtgaatac atgttataat
     1441 aactataact aataacgtaa cgtgactggc aagagatatt tttaaaacaa tgaataggtt
     1501 tacacttact ttagttttat ggaaatgaaa gatcatatca tatataatct agaataaaat
     1561 taactaaaat aattattatc tagataaaaa atttagaagc caatgaaatc tataaataaa
     1621 ctaaattaag tttatttaat taacaactat ggatataaaa taggtactaa tcaaaatagt
     1681 gaggaggata tatttgaata catacgaaca aattaataaa gtgaaaaaaa tacttcggaa
     1741 acatttaaaa aataacctta ttggtactta catgtttgga tcaggagttg agagtggact
     1801 aaaaccaaat agtgatcttg actttttagt cgtcgtatct gaaccattga cagatcaaag
     1861 taaagaaata cttatacaaa aaattagacc tatttcaaaa aaaataggag ataaaagcaa
     1921 cttacgatat attgaattaa caattattat tcagcaagaa atggtaccgt ggaatcatcc
     1981 tcccaaacaa gaatttattt atggagaatg gttacaagag ctttatgaac aaggatacat
     2041 tcctcagaag gaattaaatt cagatttaac cataatgctt taccaagcaa aacgaaaaaa
     2101 taaaagaata tacggaaatt atgacttaga ggaattacta cctgatattc cattttctga
     2161 tgtgagaaga gccattatgg attcgtcaga ggaattaata gataattatc aggatgatga
     2221 aaccaactct atattaactt tatgccgtat gattttaact atggacacgg gtaaaatcat
     2281 accaaaagat attgcgggaa atgcagtggc tgaatcttct ccattagaac atagggagag
     2341 aattttgtta gcagttcgta gttatcttgg agagaatatt gaatggacta atgaaaatgt
     2401 aaatttaact ataaactatt taaataacag attaaaaaaa ttataaaaaa attgaaaaaa
     2461 tggtggaaac acttttttca atttttttgt tttattattt aatatttggg aaatattcat
     2521 tctaattggt aatcagattt tagaaaacaa taaacccttg cataactttc tcgtccatat
     2581 cggaaactac ttttttattc cctttttttc gaccgagatt attttgcgat aaaatccgat
     2641 taagatactg cctactaact cccaattctt gagctaattc tgatactgtt ttactcatca
     2701 gctacctcac aaaataattc tttctatccc aattccaaaa ggcaacaatt tcacgaccag
     2761 tactttctgc agattcttct gctccggtct gaacaagatt cccttcttcg acatcatcaa
     2821 gagctaactc cgccttgatt ttcttgaaca actttccaaa accaatctgc ctttttcggt
     2881 gcaaaccatt cgttaaatct tctgttatcc gttctttgac catttcagag aataatgtca
     2941 catcttctgt atctacatca aaaggcttaa caggatattt agctgtttcg ataatagctg
     3001 acttcaagtt tttacgttta tgagctttga cggttcgaat atcaactaca ggtgtataat
     3061 ccaatttcat cgccttagcc catagtttcg tccattcttc ttggctaatg taatttgtat
     3121 tatttccagt aaaatagcta gatttcacaa ataacaatac atgcaaatga gggtgataat
     3181 tctcatgctc agttgaataa gttacctcag tggctcgcaa atagccaata acatttttat
     3241 cgactttctt gtatttcata agacgattaa aagctcgccc tatttctgaa attgatttat
     3301 ttaatttttc accactaatg ttcttaatcg tcaaggtcaa aaagagaaag cgaccttttg
     3361 gatagcgttt cattgcttca tttaccacta attcagcttg ataagcatac ttcattgacc
     3421 gtctccagtt acaaattggg cagagcttat ttttgcaaaa ataagactga tagagttttt
     3481 tcgtgccgtc ttgttgttcg ataaacttca acacttcggc acattgatag acccgttcaa
     3541 acgaacgata tcctaaacgc tctaaacgac cagcaagctc aatatttttt aacttgcgtt
     3601 ctcgccactt tctatctttg ccattcttag aatagtcctt aaagacttga ttttcagtac
     3661 ctgtcatgtt ataattttct tgcaaaaaaa tttttttgag tatataaaaa agttcctgca
     3721 attctcctaa gtttattatt ttaaattgga cacttaaatt ataaacttta ttttattaaa
     3781 ttgcaagggc ttttttgtta cctaaaaacc cagcaatatc aagggtttga ggtgctttaa
     3841 aacaagaaaa taaaaaaact cattatttct atatgtatca agataagaaa aaacccttgc
     3901 tacgcatggg tttttaaggg cttttcagcc ctctttttcg tttcacgaaa aattcacccc
     3961 taaaacctca aattttgact ttctaaatcc aaacgtggta taataatttt aacgctagaa
     4021 gttctaggaa acgaatagca caacgcaaaa aaaacagtag ctggaactac tgtctttctt
     4081 ttttgtcctc aaaaagaact caggttaatt tttgtcgcgt ttgtttcgaa cataccgaat
     4141 caaccaatca gcacttacct ctgcagcaac gccaaccaca aaaggtatca aaacaagttc
     4201 taggaaagtc ttcacagcgc tcacctccaa tctaagttcg taacttagcg aaagaacgtt
     4261 gcgacctagc tattatacca tgtccgctta ctttattcca tggtctaatt gacaacaagt
     4321 aaccaagggc ggacgtcctt tgtccgtgtc ggctcgaaac gctaaagcct ttcggctcgt
     4381 cacgtcctag cgtactttgc ccttgcttat tgtcaattag ctttcacggc ataaatcgct
     4441 caaaggccta gccctttttc aatcactcgt ttaactaccc ttacgattgg ctgaatagct
     4501 cttaactctg atataaaatt ttttaaggct ttatctggca taaactcttt cagttttcct
     4561 agcattttgt cattttgctg tctcaaaact gagttttcac ttctaagcgc tctattttcg
     4621 ttcctaagac gctctatgct ctctccctta cttttttctt tagctcgttc caaactcgct
     4681 tcaaaaccgt ttttagagcg tctcagaggc actatccgcc ttaattgctc attctctagc
     4741 tctttttgat gaacaaggtt tctgcttcgg tatatcagcc ctttaaattc ttcaaactgc
     4801 tctttagtca attccacata ttctttaccg aacacgcctt ttttaggggt aatagccgct
     4861 agttctttct cagacatgat ttttaggtca gctatttgtg tttttagttt ttctaatttc
     4921 agcccctcgc gctctaaacg ctctattttc gcttctaagc tcttcgaata attttctaag
     4981 gtatttatat atccctcagc tttagatagc tctgagagcc tctcagaggt ctcagaatcc
     5041 caacgctgcc cgactgcttt cctgatgcaa aaacgaggct agtttaccgt atctgtgggg
     5101 ggatggcttg tagatatgac gacaggaaga gtttgtagaa acgcaaaaag gccatccgtc
     5161 aggatggcct tctgcttaat ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     5221 cctccgggcc gttgcttcgc aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     5281 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     5341 cgttttattt gatgcctggc agttccctac tctcgcatgg ggagacccca cactaccatc
     5401 ggcgctacgg cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact
     5461 gccgccaggc aaattctgtt ttatcagacc gcttctgcgt tctgatttaa tctgtatcag
     5521 gctgaaaatc ttctctcatc cgccaaaaca gctgctttcc tgatgcaaaa acgaggctag
     5581 tttaccgtat ctgtgggggg atggcttgta gatatgacga caggaagagt ttgtagaaac
     5641 gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc agtttatggc
     5701 gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc cgctcccggc
     5761 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     5821 tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg
     5881 agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg
     5941 tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc ttctgcgttc
     6001 tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc cgagatgcgc
     6061 cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc
     6121 gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt gaataattcc
     6181 cggcattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6241 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6301 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg
     6361 ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     6421 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6481 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6541 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6601 ggggagaggc ggtttgcgta ttggagcttg ttgtaactga aaaaggaaaa ttattgtgcc
     6661 aggcagttga aagtcagcac cttttaacga gtgctgaaat gacggctaaa tgggaaacgt
     6721 attt
//
