LOCUS       pBAMD1-4        4730 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Standardized mini-Tn5 delivery plasmid, streptomycin/spectinomycin 
            resistant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4730)
  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 4730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
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: pBAMD1-4
FEATURES             Location/Qualifiers
     source          1..4730
                     /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"
     CDS             362..1153
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    1246..1264
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     terminator      1275..1369
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1474..2901
                     /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        2958..3049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3050..3118,3119..3910)
                     /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      3914..3953
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            4101..4209
                     /note="incP origin of transfer"
     rep_origin      4220..4608
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      complement(4639..4725)
                     /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 aaatgaacct tgaccgaacg
      181 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
      241 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
      301 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
      361 catgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
      421 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
      481 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
      541 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
      601 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
      661 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
      721 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
      781 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
      841 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
      901 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
      961 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     1021 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     1081 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
     1141 agtcggcaaa taagacaatt gtctaattaa ttgcggaccc tagaggtccc cttttttatt
     1201 ttaaaaattt tttcacaaaa cggtttacaa gcataaaatc tctgaagatg tgtataagag
     1261 acagactagt cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     1321 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc
     1381 ctggtttaaa ctacacaagt agcgtcctga acggaacctt tcccgttttc cagaatctga
     1441 tgttccatgt gacctcctaa catggtaacg ttcatgatta ccagtgcact gcatcgtgcg
     1501 gcggattggg cgaaaagcgt gttttctagt gctgcgctgg gtgatccgcg tcgtaccgcg
     1561 cgtctggtga atgttgcggc gcaactggcc aaatatagcg gcaaaagcat taccattagc
     1621 agcgaaggca gcaaagccat gcaggaaggc gcgtatcgtt ttattcgtaa tccgaacgtg
     1681 agcgcggaag cgattcgtaa agcgggtgcc atgcagaccg tgaaactggc ccaggaattt
     1741 ccggaactgc tggcaattga agataccacc tctctgagct atcgtcatca ggtggcggaa
     1801 gaactgggca aactgggtag cattcaggat aaaagccgtg gttggtgggt gcatagcgtg
     1861 ctgctgctgg aagcgaccac ctttcgtacc gtgggcctgc tgcatcaaga atggtggatg
     1921 cgtccggatg atccggcgga tgcggatgaa aaagaaagcg gcaaatggct ggccgctgct
     1981 gcaacttcgc gtctgagaat gggcagcatg atgagcaacg tgattgcggt gtgcgatcgt
     2041 gaagcggata ttcatgcgta tctgcaagat aaactggccc ataacgaacg ttttgtggtg
     2101 cgtagcaaac atccgcgtaa agatgtggaa agcggcctgt atctgtatga tcacctgaaa
     2161 aaccagccgg aactgggcgg ctatcagatt agcattccgc agaaaggcgt ggtggataaa
     2221 cgtggcaaac gtaaaaaccg tccggcgcgt aaagcgagcc tgagcctgcg tagcggccgt
     2281 attaccctga aacagggcaa cattaccctg aacgcggtgc tggccgaaga aattaatccg
     2341 ccgaaaggcg aaaccccgct gaaatggctg ctgctgacca gcgagccggt ggaaagtctg
     2401 gcccaagcgc tgcgtgtgat tgatatttat acccatcgtt ggcgcattga agaatttcac
     2461 aaagcgtgga aaacgggtgc gggtgcggaa cgtcagcgta tggaagaacc ggataacctg
     2521 gaacgtatgg tgagcattct gagctttgtg gcggtgcgtc tgctgcaact gcgtgaatct
     2581 tttactccgc cgcaagcact gcgtgcgcag ggcctgctga aagaagcgga acacgttgaa
     2641 agccagagcg cggaaaccgt gctgaccccg gatgaatgcc aactgctggg ctatctggat
     2701 aaaggcaaac gcaaacgcaa agaaaaagcg ggcagcctgc aatgggcgta tatggcgatt
     2761 gcgcgtctgg gcggctttat ggatagcaaa cgtaccggca ttgcgagctg gggtgcgctg
     2821 tgggaaggtt gggaagcgct gcaaagcaaa ctggatggct ttctggccgc gaaagacctg
     2881 atggcgcagg gcattaaaat ctaatcggcg atcgctagta gcccgcctaa tgagcgggct
     2941 tttttttaat tcccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3001 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagcattca
     3061 gcattttcgt gtggcgctga ttccgttttt tgcggcgttt tgcctgccgg tgtttgcgca
     3121 tccggaaacc ctggtgaaag tgaaagatgc ggaagatcaa ctgggtgcgc gcgtgggcta
     3181 tattgaactg gatctgaaca gcggcaaaat tctggaatct tttcgtccgg aagaacgttt
     3241 tccgatgatg agcaccttta aagtgctgct gtgcggtgcg gttctgagcc gtgtggatgc
     3301 gggccaggaa caactgggcc gtcgtattca ttatagccag aacgatctgg tggaatatag
     3361 cccggtgacc gaaaaacatc tgaccgatgg catgaccgtg cgtgaactgt gcagcgcggc
     3421 gattaccatg agcgataaca ccgcggcgaa cctgctgctg acgaccattg gcggtccgaa
     3481 agaactgacc gcgtttctgc ataacatggg cgatcatgtg acccgtctgg atcgttggga
     3541 accggaactg aacgaagcga ttccgaacga tgaacgtgat accaccatgc cggcagcaat
     3601 ggcgaccacc ctgcgtaaac tgctgacggg tgagctgctg accctggcaa gccgccagca
     3661 actgattgat tggatggaag cggataaagt ggcgggtccg ctgctgcgta gcgcgctgcc
     3721 ggctggctgg tttattgcgg ataaaagcgg tgcgggcgaa cgtggcagcc gtggcattat
     3781 tgcggcgctg ggcccggatg gtaaaccgag ccgtattgtg gtgatttata ccaccggcag
     3841 ccaggcgacg atggatgaac gtaaccgtca gattgcggaa attggcgcga gcctgattaa
     3901 acattggtaa accgatacaa ttaaaggctc cttttggagc cttttttttt ggacacgcgt
     3961 ttgtcctttt ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat
     4021 ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg
     4081 tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt
     4141 tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga
     4201 ggctggccgt aggccggccg atctgaagat cagcagttca acctgttgat agtacgtact
     4261 aagctctcat gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa
     4321 cgaactaaac cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat
     4381 gtttcacgta ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa
     4441 tagcctctaa ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagcct
     4501 ttaaggttta acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga
     4561 gcctctcaaa gcaattttga gtgacacagg aacacttaac ggctgacatg gggcgcgccc
     4621 agctgtctag ggcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     4681 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc
//