LOCUS       pVAM05                  5582 bp    DNA     circular SYN 03-JUN-2024
DEFINITION  pVAM05.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5582)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 from SnapGene 8.1.1
            https://www.snapgene.com
COMMENT     SECID/File created by Clone Manager, Scientific
FEATURES             Location/Qualifiers
     source          1..5582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..392
                     /gene="R6k_origin"
                     /label=R6k_origin
     misc_feature    1461..1559
                     /gene="oriT_RP4"
                     /label=oriT_RP4
     CDS             2576..3109
                     /codon_start=1
                     /gene="gentamicin"
                     /product="gentamicin-3-acetyltransferase"
                     /label=gentamicin
                     /note="gentamicin-3-acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             complement(3140..4561)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial
                     growth sensitive to sucrose"
                     /label=Bacillus subtilis sacB
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             5241..5582
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ fragment
                     /label=lacZ-alpha
                     /translation="MTMITAAANSSSVPGDPLESTCRHASLGPALAVVLQRRDWENPGV
                     TQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIRQSN
                     HSTRPVAAH"
     misc_feature    5264..5320
                     /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 acataatgtt
     2581 acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggtggctc
     2641 aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat ccatgcgggc
     2701 tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc aacatcagcc
     2761 ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt
     2821 cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaggt ttgagcagcc
     2881 gcgtagtgag atctatatct atgatctcgc agtctccggc gagcaccgga ggcagggcat
     2941 tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg cttatgtgat
     3001 ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa agttgggcat
     3061 acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcccatc
     3121 ggcattttct tttgcgtttt tatttgttaa ctgttaattg tccttgttca aggatgctgt
     3181 ctttgacaac agatgttttc ttgcctttga tgttcagcag gaagcttggc gcaaacgttg
     3241 attgtttgtc tgcgtagaat cctctgtttg tcatatagct tgtaatcacg acattgtttc
     3301 ctttcgcttg aggtacagcg aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat
     3361 ccatttttaa cacaaggcca gttttgttca gcggcttgta tgggccagtt aaagaattag
     3421 aaacataacc aagcatgtaa atatcgttag acgtaatgcc gtcaatcgtc atttttgatc
     3481 cgcgggagtc agtgaacagg taccatttgc cgttcatttt aaagacgttc gcgcgttcaa
     3541 tttcatctgt tactgtgtta gatgcaatca gcggtttcat cacttttttc agtgtgtaat
     3601 catcgtttag ctcaatcata ccgagagcgc cgtttgctaa ctcagccgtg cgttttttat
     3661 cgctttgcag aagtttttga ctttcttgac ggaagaatga tgtgcttttg ccatagtatg
     3721 ctttgttaaa taaagattct tcgccttggt agccatcttc agttccagtg tttgcttcaa
     3781 atactaagta tttgtggcct ttatcttcta cgtagtgagg atctctcagc gtatggttgt
     3841 cgcctgagct gtagttgcct tcatcgatga actgctgtac attttgatac gtttttccgt
     3901 caccgtcaaa gattgattta taatcctcta caccgttgat gttcaaagag ctgtctgatg
     3961 ctgatacgtt aacttgtgca gttgtcagtg tttgtttgcc gtaatgttta ccggagaaat
     4021 cagtgtagaa taaacggatt tttccgtcag atgtaaatgt ggctgaacct gaccattctt
     4081 gtgtttggtc ttttaggata gaatcatttg catcgaattt gtcgctgtct ttaaagacgc
     4141 ggccagcgtt tttccagctg tcaatagaag tttcgccgac tttttgatag aacatgtaaa
     4201 tcgatgtgtc atccgcattt ttaggatctc cggctaatgc aaagacgatg tggtagccgt
     4261 gatagtttgc gacagtgccg tcagcgtttt gtaatggcca gctgtcccaa acgtccaggc
     4321 cttttgcaga agagatattt ttaattgtgg acgaatcgaa ttcaggaact tgatattttt
     4381 catttttttg ctgttcaggg atttgcagca tatcatggcg tgtaatatgg gaaatgccgt
     4441 atgtttcctt atatggcttt tggttcgttt ctttcgcaaa cgcttgagtt gcgcctcctg
     4501 ccagcagtgc ggtagtaaag gttaatactg ttgcttgttt tgcaaacttt ttgatgttca
     4561 tcgttcatgt ctcctttttt atgtactgtg ttagcggtct gcttcttcca gccctcctgt
     4621 ttgaagatgg caagttagtt acgcacaata aaaaaagacc taaaatatgt aaggggtgac
     4681 gccaaagtat acactttgcc ctttacacat tttaggtctt gcctgcttta tcagtaacaa
     4741 acccgcgcga tttacttttc gacctcattc tattagactc tcgtttggat tgcaactggt
     4801 ctattttcct cttttgtttg atagaaaatc ataaaaggat ttgcagacta cgggcctaaa
     4861 gaactaaaaa atctatctgt ttcttttcat tctctgtatt ttttatagtt tctgttgcat
     4921 gggcataaag ttgccttttt aatcacaatt cagaaaatat cataatatct catttcacta
     4981 aataatagtg aacggcaggt atatgtgatg ggttaaaaag gatcagcgcc caatacgcaa
     5041 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5101 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5161 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5221 atttcacaca ggaaacagct atgaccatga ttactgcggc cgcgaattcg agctcggtac
     5281 ccggggatcc tctagagtcg acctgcaggc atgcaagctt gggcccggca ctggccgtcg
     5341 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5401 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5461 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     5521 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     5581 aa
//
