LOCUS       sequence-266683-        4658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Standardized mini-Tn5 delivery plasmid, kanamycin resistant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4658)
  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 4658)
  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: pBAMD1-2
FEATURES             Location/Qualifiers
     source          1..4658
                     /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"
     primer_bind     complement(255..274)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             266..1081
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    1174..1192
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     terminator      1203..1297
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1402..2829
                     /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        2886..2977
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2978..3046,3047..3838)
                     /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      3842..3881
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            4029..4137
                     /note="incP origin of transfer"
     rep_origin      4148..4536
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      complement(4567..4653)
                     /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 aaatttgtgt ctcaaaatct
      181 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
      241 cataaacagt aatacaaggg gtgttatgag ccatattcag cgtgaaacga gctgtagccg
      301 tccgcgtctg aacagcaaca tggatgcgga tctgtatggc tataaatggg cgcgtgataa
      361 cgtgggtcag agcggcgcga ccatttatcg tctgtatggc aaaccggatg cgccggaact
      421 gtttctgaaa catggcaaag gcagcgtggc gaacgatgtg accgatgaaa tggtgcgtct
      481 gaactggctg accgaattta tgccgctgcc gaccattaaa cattttattc gcaccccgga
      541 tgatgcgtgg ctgctgacca ccgcgattcc gggcaaaacc gcgtttcagg tgctggaaga
      601 atatccggat agcggcgaaa acattgtgga tgcgctggcc gtgtttctgc gtcgtctgca
      661 tagcattccg gtgtgcaact gcccgtttaa cagcgatcgt gtgtttcgtc tggcccaggc
      721 gcagagccgt atgaacaacg gcctggtgga tgcgagcgat tttgatgatg aacgtaacgg
      781 ctggccggtg gaacaggtgt ggaaagaaat gcataaactg ctgccgttta gcccggatag
      841 cgtggtgacc cacggcgatt ttagcctgga taacctgatt ttcgatgaag gcaaactgat
      901 tggctgcatt gatgtgggcc gtgtgggcat tgcggatcgt tatcaggatc tggccattct
      961 gtggaactgc ctgggcgaat ttagcccgag cctgcaaaaa cgtctgtttc agaaatatgg
     1021 cattgataat ccggatatga acaaactgca atttcatctg atgctggatg aatttttcta
     1081 agacccttgt ctaattaatt gcggacccta gaggtcccct tttttatttt aaaaattttt
     1141 tcacaaaacg gtttacaagc ataaaatctc tgaagatgtg tataagagac agactagtct
     1201 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     1261 gctcggttgc cgccgggcgt tttttattgg tgagaatcca ggggtcccct ggtttaaact
     1321 acacaagtag cgtcctgaac ggaacctttc ccgttttcca gaatctgatg ttccatgtga
     1381 cctcctaaca tggtaacgtt catgattacc agtgcactgc atcgtgcggc ggattgggcg
     1441 aaaagcgtgt tttctagtgc tgcgctgggt gatccgcgtc gtaccgcgcg tctggtgaat
     1501 gttgcggcgc aactggccaa atatagcggc aaaagcatta ccattagcag cgaaggcagc
     1561 aaagccatgc aggaaggcgc gtatcgtttt attcgtaatc cgaacgtgag cgcggaagcg
     1621 attcgtaaag cgggtgccat gcagaccgtg aaactggccc aggaatttcc ggaactgctg
     1681 gcaattgaag ataccacctc tctgagctat cgtcatcagg tggcggaaga actgggcaaa
     1741 ctgggtagca ttcaggataa aagccgtggt tggtgggtgc atagcgtgct gctgctggaa
     1801 gcgaccacct ttcgtaccgt gggcctgctg catcaagaat ggtggatgcg tccggatgat
     1861 ccggcggatg cggatgaaaa agaaagcggc aaatggctgg ccgctgctgc aacttcgcgt
     1921 ctgagaatgg gcagcatgat gagcaacgtg attgcggtgt gcgatcgtga agcggatatt
     1981 catgcgtatc tgcaagataa actggcccat aacgaacgtt ttgtggtgcg tagcaaacat
     2041 ccgcgtaaag atgtggaaag cggcctgtat ctgtatgatc acctgaaaaa ccagccggaa
     2101 ctgggcggct atcagattag cattccgcag aaaggcgtgg tggataaacg tggcaaacgt
     2161 aaaaaccgtc cggcgcgtaa agcgagcctg agcctgcgta gcggccgtat taccctgaaa
     2221 cagggcaaca ttaccctgaa cgcggtgctg gccgaagaaa ttaatccgcc gaaaggcgaa
     2281 accccgctga aatggctgct gctgaccagc gagccggtgg aaagtctggc ccaagcgctg
     2341 cgtgtgattg atatttatac ccatcgttgg cgcattgaag aatttcacaa agcgtggaaa
     2401 acgggtgcgg gtgcggaacg tcagcgtatg gaagaaccgg ataacctgga acgtatggtg
     2461 agcattctga gctttgtggc ggtgcgtctg ctgcaactgc gtgaatcttt tactccgccg
     2521 caagcactgc gtgcgcaggg cctgctgaaa gaagcggaac acgttgaaag ccagagcgcg
     2581 gaaaccgtgc tgaccccgga tgaatgccaa ctgctgggct atctggataa aggcaaacgc
     2641 aaacgcaaag aaaaagcggg cagcctgcaa tgggcgtata tggcgattgc gcgtctgggc
     2701 ggctttatgg atagcaaacg taccggcatt gcgagctggg gtgcgctgtg ggaaggttgg
     2761 gaagcgctgc aaagcaaact ggatggcttt ctggccgcga aagacctgat ggcgcagggc
     2821 attaaaatct aatcggcgat cgctagtagc ccgcctaatg agcgggcttt tttttaattc
     2881 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2941 ctgataaatg cttcaataat attgaaaaag gaagagtatg agcattcagc attttcgtgt
     3001 ggcgctgatt ccgttttttg cggcgttttg cctgccggtg tttgcgcatc cggaaaccct
     3061 ggtgaaagtg aaagatgcgg aagatcaact gggtgcgcgc gtgggctata ttgaactgga
     3121 tctgaacagc ggcaaaattc tggaatcttt tcgtccggaa gaacgttttc cgatgatgag
     3181 cacctttaaa gtgctgctgt gcggtgcggt tctgagccgt gtggatgcgg gccaggaaca
     3241 actgggccgt cgtattcatt atagccagaa cgatctggtg gaatatagcc cggtgaccga
     3301 aaaacatctg accgatggca tgaccgtgcg tgaactgtgc agcgcggcga ttaccatgag
     3361 cgataacacc gcggcgaacc tgctgctgac gaccattggc ggtccgaaag aactgaccgc
     3421 gtttctgcat aacatgggcg atcatgtgac ccgtctggat cgttgggaac cggaactgaa
     3481 cgaagcgatt ccgaacgatg aacgtgatac caccatgccg gcagcaatgg cgaccaccct
     3541 gcgtaaactg ctgacgggtg agctgctgac cctggcaagc cgccagcaac tgattgattg
     3601 gatggaagcg gataaagtgg cgggtccgct gctgcgtagc gcgctgccgg ctggctggtt
     3661 tattgcggat aaaagcggtg cgggcgaacg tggcagccgt ggcattattg cggcgctggg
     3721 cccggatggt aaaccgagcc gtattgtggt gatttatacc accggcagcc aggcgacgat
     3781 ggatgaacgt aaccgtcaga ttgcggaaat tggcgcgagc ctgattaaac attggtaaac
     3841 cgatacaatt aaaggctcct tttggagcct ttttttttgg acacgcgttt gtccttttcc
     3901 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
     3961 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
     4021 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
     4081 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag
     4141 gccggccgat ctgaagatca gcagttcaac ctgttgatag tacgtactaa gctctcatgt
     4201 ttcacgtact aagctctcat gtttaacgta ctaagctctc atgtttaacg aactaaaccc
     4261 tcatggctaa cgtactaagc tctcatggct aacgtactaa gctctcatgt ttcacgtact
     4321 aagctctcat gtttgaacaa taaaattaat ataaatcagc aacttaaata gcctctaagg
     4381 ttttaagttt tataagaaaa aaaagaatat ataaggcttt taaagccttt aaggtttaac
     4441 ggttgtggac aacaagccag ggatgtaacg cactgagaag cccttagagc ctctcaaagc
     4501 aattttgagt gacacaggaa cacttaacgg ctgacatggg gcgcgcccag ctgtctaggg
     4561 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     4621 ccagtctttc gactgagcct ttcgttttat ttgatgcc
//
