LOCUS       pVAM03                  6240 bp    DNA     circular SYN 03-JUN-2024
DEFINITION  pVAM03
ACCESSION   pVAM03
KEYWORDS    .
SOURCE      Unknown.
  ORGANISM  Unknown
            Unclassified.
REFERENCE   1  (bases 1 to 6240)
  AUTHORS   Self
  JOURNAL   Unpublished.
COMMENT     SECID/File created by Clone Manager, Scientific & Educational Software
FEATURES             Location/Qualifiers
     misc_feature    1..392
                     /gene="R6Kg_origin"
     misc_feature    1461..1559
                     /gene="oriT_RP4"
     CDS             2575..3765
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASH
                     YGVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAG
                     RIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLH
                     APFLAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTV
                     FFIMQLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGE
                     KQAIIAGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQG
                     QLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSR
                     ATST"
     CDS             complement(3798..5219)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGIS
                     HITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHG
                     YHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQT
                     QEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIF
                     DGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEES
                     LFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIAS
                     NTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLN
                     KTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLN
                     IKGKKTSVVKDSILEQGQLTVNK"
     CDS             5899..6240
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITAAANSSSVPGDPLESTCRHASLGPALAVVLQRRDWENPG
                     VTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIRQ
                     SNHSTRPVAAH"
     misc_feature    5922..5978
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN      
        1 ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc
       61 ttctcagtgc gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa
      121 aagccttata tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg
      181 ctgatttata ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct
      241 tagtacgtta gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg
      301 agagcttagt acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac
      361 tatcaacagg ttgaactgct gatcttcaga tcctctacgc cggacgcatc gtggccggat
      421 ccagccgacc aggctttcca cgcccgcgtg ccgctccatg tcgttcgcgc ggttctcgga
      481 aacgcgctgc cgcgtttcgt gattgtcacg ctcaagcccg tagtcccgtt cgagcgtcgc
      541 gcagaggtca gcgagggcgc ggtaggcccg atacggctca tggatggtgt ttcgggtcgg
      601 gtgaatcttg ttgatggcga tatggatgtg caggttgtcg gtgtcgtgat gcacggcact
      661 gacgcgctga tgctcggcga agccaagccc agcgcagatg cggtcctcaa tcgcgcgcaa
      721 cgtctccgcg tcgggcttct ctcccgcgcg gaagctaacc agcaggtgat aggtcttgtc
      781 ggcctcggaa cgggtgttgc cgtgctgggt cgccatcacc tcggccatga cagcgggcag
      841 ggtgtttgcc tcgcagttcg tgacgcgcac gtgacccagg cgctcggtct tgccttgctc
      901 gtcggtgatg tacttcacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac
      961 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg
     1021 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct
     1081 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
     1141 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
     1201 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
     1261 aggccgacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
     1321 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     1381 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     1441 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     1501 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     1561 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     1621 ctataatgac cccgaagcag ggttatgcag cggaaaagcg ctgcttccct gctgttttgt
     1681 ggaatatcta ccgactggaa acaggcaaat gcaggaaatt actgaactga ggggacaggc
     1741 gagagacgat gccaaagagc tacaccgacg agctggccga gtgggttgaa tcccgcgcgg
     1801 ccaagaagcg ccggcgtgat gaggctgcgg ttgcgttcct ggcggtgagg gcggatgtcg
     1861 aggcggcgtt agcgtccggc tatgcgctcg tcaccatttg ggagcacatg cgggaaacgg
     1921 ggaaggtcaa gttctcctac gagacgttcc gctcgcacgc caggcggcac atcaaggcca
     1981 agcccgccga tgtgcccgca ccgcaggcca aggctgcgga acccgcgccg gcacccaaga
     2041 cgccggagcc acggcggccg aagcaggggg gcaaggctga aaagccggcc cccgctgcgg
     2101 ccccgaccgg cttcaccttc aacccaacac cggacaaaaa ggatcctcta cgccggacgc
     2161 atcgtggccg gcatcaccgg cgccacaagt gcggttgctg gcgcctatat cgccgacatc
     2221 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     2281 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgct gcctcgcgcg
     2341 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2401 tctgtaagcg gtttgggttg tcactcatcg tatttggttt ataattttaa gctaataacc
     2461 taattataac taattaatag ttttgtatct tgaaaaaata gctataaaac ttatttaaat
     2521 aacgaagatt tttgtgtata aaatatttat aacaaaaaaa aggagactaa acatatgaaa
     2581 tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc
     2641 ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc
     2701 agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt
     2761 ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga
     2821 gccactatcg actacgcgat catggcgacc acacccgtcc tgtggatcct ctacgccgga
     2881 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     2941 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     3001 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
     3061 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     3121 gagtcgcata agggagagcg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     3181 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     3241 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     3301 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     3361 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     3421 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg
     3481 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     3541 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     3601 cttaccagcc taacttcgat cactggaccg ctgatcgtca cggcgattta tgccgcctcg
     3661 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc
     3721 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga agcccatcgg
     3781 cattttcttt tgcgttttta tttgttaact gttaattgtc cttgttcaag gatgctgtct
     3841 ttgacaacag atgttttctt gcctttgatg ttcagcagga agcttggcgc aaacgttgat
     3901 tgtttgtctg cgtagaatcc tctgtttgtc atatagcttg taatcacgac attgtttcct
     3961 ttcgcttgag gtacagcgaa gtgtgagtaa gtaaaggtta catcgttagg atcaagatcc
     4021 atttttaaca caaggccagt tttgttcagc ggcttgtatg ggccagttaa agaattagaa
     4081 acataaccaa gcatgtaaat atcgttagac gtaatgccgt caatcgtcat ttttgatccg
     4141 cgggagtcag tgaacaggta ccatttgccg ttcattttaa agacgttcgc gcgttcaatt
     4201 tcatctgtta ctgtgttaga tgcaatcagc ggtttcatca cttttttcag tgtgtaatca
     4261 tcgtttagct caatcatacc gagagcgccg tttgctaact cagccgtgcg ttttttatcg
     4321 ctttgcagaa gtttttgact ttcttgacgg aagaatgatg tgcttttgcc atagtatgct
     4381 ttgttaaata aagattcttc gccttggtag ccatcttcag ttccagtgtt tgcttcaaat
     4441 actaagtatt tgtggccttt atcttctacg tagtgaggat ctctcagcgt atggttgtcg
     4501 cctgagctgt agttgccttc atcgatgaac tgctgtacat tttgatacgt ttttccgtca
     4561 ccgtcaaaga ttgatttata atcctctaca ccgttgatgt tcaaagagct gtctgatgct
     4621 gatacgttaa cttgtgcagt tgtcagtgtt tgtttgccgt aatgtttacc ggagaaatca
     4681 gtgtagaata aacggatttt tccgtcagat gtaaatgtgg ctgaacctga ccattcttgt
     4741 gtttggtctt ttaggataga atcatttgca tcgaatttgt cgctgtcttt aaagacgcgg
     4801 ccagcgtttt tccagctgtc aatagaagtt tcgccgactt tttgatagaa catgtaaatc
     4861 gatgtgtcat ccgcattttt aggatctccg gctaatgcaa agacgatgtg gtagccgtga
     4921 tagtttgcga cagtgccgtc agcgttttgt aatggccagc tgtcccaaac gtccaggcct
     4981 tttgcagaag agatattttt aattgtggac gaatcgaatt caggaacttg atatttttca
     5041 tttttttgct gttcagggat ttgcagcata tcatggcgtg taatatggga aatgccgtat
     5101 gtttccttat atggcttttg gttcgtttct ttcgcaaacg cttgagttgc gcctcctgcc
     5161 agcagtgcgg tagtaaaggt taatactgtt gcttgttttg caaacttttt gatgttcatc
     5221 gttcatgtct ccttttttat gtactgtgtt agcggtctgc ttcttccagc cctcctgttt
     5281 gaagatggca agttagttac gcacaataaa aaaagaccta aaatatgtaa ggggtgacgc
     5341 caaagtatac actttgccct ttacacattt taggtcttgc ctgctttatc agtaacaaac
     5401 ccgcgcgatt tacttttcga cctcattcta ttagactctc gtttggattg caactggtct
     5461 attttcctct tttgtttgat agaaaatcat aaaaggattt gcagactacg ggcctaaaga
     5521 actaaaaaat ctatctgttt cttttcattc tctgtatttt ttatagtttc tgttgcatgg
     5581 gcataaagtt gcctttttaa tcacaattca gaaaatatca taatatctca tttcactaaa
     5641 taatagtgaa cggcaggtat atgtgatggg ttaaaaagga tcagcgccca atacgcaaac
     5701 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5761 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     5821 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     5881 ttcacacagg aaacagctat gaccatgatt actgcggccg cgaattcgag ctcggtaccc
     5941 ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg gcccggcact ggccgtcgtt
     6001 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     6061 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6121 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     6181 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
//
