LOCUS       sequence-100351-        4546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Standardized mini-Tn5 delivery plasmid, gentamicin resistant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4546)
  AUTHORS   Martinez-Garcia E, Aparicio T, de Lorenzo V, Nikel PI
  TITLE     New transposon tools tailored for metabolic engineering of 
            gram-negative microbial cell factories.
  JOURNAL   Front Bioeng Biotechnol. 2014 Oct 28;2:46. doi: 
            10.3389/fbioe.2014.00046. eCollection 2014.
  PUBMED    25389526
REFERENCE   2  (bases 1 to 4546)
  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: "Front 
            Bioeng Biotechnol."; date: "2014-10-28"; volume: "2:46. doi"; pages:
            " 10.3389/fbioe.2014.00046. eCollection 2014"
COMMENT     Sequence Label: pBAMD1-6
FEATURES             Location/Qualifiers
     source          1..4546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..28
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     misc_feature    62..118
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        join(165..170,171..187,188..193)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             382..915
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     misc_feature    1062..1080
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     terminator      1091..1185
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1290..2717
                     /codon_start=1
                     /product="transposase from the bacterial Tn5 transposon 
                     (Reznikoff, 1993)"
                     /label=Tn5 transposase
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     promoter        2774..2865
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2866..2934,2935..3726)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3730..3769
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            3917..4025
                     /note="incP origin of transfer"
     rep_origin      4036..4424
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      complement(4455..4541)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tttaattaac tgtctcttat acacatcttt gtgtctcagg ccgcctaggc cgcggccgcg
       61 cgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgc
      121 ggccgccaaa gcccgccgaa aggcgggctt ttctgtattt aaatttgaca taagcctgtt
      181 cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
      241 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
      301 ttgtacagcc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
      361 tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt
      421 cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc
      481 cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac
      541 gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt
      601 aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct
      661 tacgttctgc ccaagtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc
      721 tccggagagc accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag
      781 gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca
      841 gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca
      901 agtaccgcca cctaacaatt cgttcaagcc gagatcggct tcccggccgc ggagttgttc
      961 ggtaaattgg acaattgtct aattaattgc ggaccctaga ggtccccttt tttattttaa
     1021 aaattttttc acaaaacggt ttacaagcat aaaatctctg aagatgtgta taagagacag
     1081 actagtcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     1141 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccagg ggtcccctgg
     1201 tttaaactac acaagtagcg tcctgaacgg aacctttccc gttttccaga atctgatgtt
     1261 ccatgtgacc tcctaacatg gtaacgttca tgattaccag tgcactgcat cgtgcggcgg
     1321 attgggcgaa aagcgtgttt tctagtgctg cgctgggtga tccgcgtcgt accgcgcgtc
     1381 tggtgaatgt tgcggcgcaa ctggccaaat atagcggcaa aagcattacc attagcagcg
     1441 aaggcagcaa agccatgcag gaaggcgcgt atcgttttat tcgtaatccg aacgtgagcg
     1501 cggaagcgat tcgtaaagcg ggtgccatgc agaccgtgaa actggcccag gaatttccgg
     1561 aactgctggc aattgaagat accacctctc tgagctatcg tcatcaggtg gcggaagaac
     1621 tgggcaaact gggtagcatt caggataaaa gccgtggttg gtgggtgcat agcgtgctgc
     1681 tgctggaagc gaccaccttt cgtaccgtgg gcctgctgca tcaagaatgg tggatgcgtc
     1741 cggatgatcc ggcggatgcg gatgaaaaag aaagcggcaa atggctggcc gctgctgcaa
     1801 cttcgcgtct gagaatgggc agcatgatga gcaacgtgat tgcggtgtgc gatcgtgaag
     1861 cggatattca tgcgtatctg caagataaac tggcccataa cgaacgtttt gtggtgcgta
     1921 gcaaacatcc gcgtaaagat gtggaaagcg gcctgtatct gtatgatcac ctgaaaaacc
     1981 agccggaact gggcggctat cagattagca ttccgcagaa aggcgtggtg gataaacgtg
     2041 gcaaacgtaa aaaccgtccg gcgcgtaaag cgagcctgag cctgcgtagc ggccgtatta
     2101 ccctgaaaca gggcaacatt accctgaacg cggtgctggc cgaagaaatt aatccgccga
     2161 aaggcgaaac cccgctgaaa tggctgctgc tgaccagcga gccggtggaa agtctggccc
     2221 aagcgctgcg tgtgattgat atttataccc atcgttggcg cattgaagaa tttcacaaag
     2281 cgtggaaaac gggtgcgggt gcggaacgtc agcgtatgga agaaccggat aacctggaac
     2341 gtatggtgag cattctgagc tttgtggcgg tgcgtctgct gcaactgcgt gaatctttta
     2401 ctccgccgca agcactgcgt gcgcagggcc tgctgaaaga agcggaacac gttgaaagcc
     2461 agagcgcgga aaccgtgctg accccggatg aatgccaact gctgggctat ctggataaag
     2521 gcaaacgcaa acgcaaagaa aaagcgggca gcctgcaatg ggcgtatatg gcgattgcgc
     2581 gtctgggcgg ctttatggat agcaaacgta ccggcattgc gagctggggt gcgctgtggg
     2641 aaggttggga agcgctgcaa agcaaactgg atggctttct ggccgcgaaa gacctgatgg
     2701 cgcagggcat taaaatctaa tcggcgatcg ctagtagccc gcctaatgag cgggcttttt
     2761 tttaattccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2821 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag cattcagcat
     2881 tttcgtgtgg cgctgattcc gttttttgcg gcgttttgcc tgccggtgtt tgcgcatccg
     2941 gaaaccctgg tgaaagtgaa agatgcggaa gatcaactgg gtgcgcgcgt gggctatatt
     3001 gaactggatc tgaacagcgg caaaattctg gaatcttttc gtccggaaga acgttttccg
     3061 atgatgagca cctttaaagt gctgctgtgc ggtgcggttc tgagccgtgt ggatgcgggc
     3121 caggaacaac tgggccgtcg tattcattat agccagaacg atctggtgga atatagcccg
     3181 gtgaccgaaa aacatctgac cgatggcatg accgtgcgtg aactgtgcag cgcggcgatt
     3241 accatgagcg ataacaccgc ggcgaacctg ctgctgacga ccattggcgg tccgaaagaa
     3301 ctgaccgcgt ttctgcataa catgggcgat catgtgaccc gtctggatcg ttgggaaccg
     3361 gaactgaacg aagcgattcc gaacgatgaa cgtgatacca ccatgccggc agcaatggcg
     3421 accaccctgc gtaaactgct gacgggtgag ctgctgaccc tggcaagccg ccagcaactg
     3481 attgattgga tggaagcgga taaagtggcg ggtccgctgc tgcgtagcgc gctgccggct
     3541 ggctggttta ttgcggataa aagcggtgcg ggcgaacgtg gcagccgtgg cattattgcg
     3601 gcgctgggcc cggatggtaa accgagccgt attgtggtga tttataccac cggcagccag
     3661 gcgacgatgg atgaacgtaa ccgtcagatt gcggaaattg gcgcgagcct gattaaacat
     3721 tggtaaaccg atacaattaa aggctccttt tggagccttt ttttttggac acgcgtttgt
     3781 ccttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat
     3841 cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta
     3901 tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct
     3961 tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct
     4021 ggccgtaggc cggccgatct gaagatcagc agttcaacct gttgatagta cgtactaagc
     4081 tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat gtttaacgaa
     4141 ctaaaccctc atggctaacg tactaagctc tcatggctaa cgtactaagc tctcatgttt
     4201 cacgtactaa gctctcatgt ttgaacaata aaattaatat aaatcagcaa cttaaatagc
     4261 ctctaaggtt ttaagtttta taagaaaaaa aagaatatat aaggctttta aagcctttaa
     4321 ggtttaacgg ttgtggacaa caagccaggg atgtaacgca ctgagaagcc cttagagcct
     4381 ctcaaagcaa ttttgagtga cacaggaaca cttaacggct gacatggggc gcgcccagct
     4441 gtctagggcg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     4501 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcc
//
