LOCUS       sequence-439060-        7109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 integrative vector with PabstBR promoter expression cassette and
            aprR marker on Tn7 transposon; empty vector plasmid with MCS for 
            cloning - we recommend NcoI/BamHI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7109)
  AUTHORS   Bacon EE, Tran JS, Nadig N, Peters JM
  TITLE     Modular, inducible, and titratable expression systems for 
            Escherichia coli and Acinetobacter baumannii.
  JOURNAL   Microbiol Spectr. 2024 Nov 5;12(11):e0130624. doi: 
            10.1128/spectrum.01306-24. Epub 2024 Sep 20.
  PUBMED    39302127
REFERENCE   2  (bases 1 to 7109)
  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: "Microbiol 
            Spectr."; date: "2024-11-5"; volume: "12(11):e0130624. doi"; pages: 
            " 10.1128/spectrum.01306-24. Epub 2024 Sep 20"
COMMENT     Sequence Label: pJMP12042
FEATURES             Location/Qualifiers
     source          1..7109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     273..292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(511..615)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     683..701
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(739..761)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     861..880
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1294..1310
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1295..1311
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1339..1433
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1536..1622
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1673..1720
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1771..2571)
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKENSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     promoter        complement(join(2760..2765,2766..2782,2783..2788))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    2906..2953
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(3471..4553)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4515..4534
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4554..4631)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    4959..5015
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5048..5065)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5218..5304
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5396..5423
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     mobile_element  5489..5654
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            5726..5834
                     /note="incP origin of transfer"
     rep_origin      6103..6491
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(6759..7109,1..441,442..510))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
       61 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
      301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
      481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
      541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      601 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
      661 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
      721 cggtgacgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
      781 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
      841 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatgtgcg
      901 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
      961 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     1021 agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa tggatgccct
     1081 gcgtaagcag gtgtgggcgg acaataaagt cttaaactga acaaaataga tctaaactat
     1141 gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc agttatgctg
     1201 tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt ctgaagtgca
     1261 aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacggtatc gataagctag
     1321 cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     1381 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagct
     1441 agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta
     1501 aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga
     1561 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1621 atccgccgct aggagcttgc ggcccggacg atgagctcga attggggatc ttgaagtacc
     1681 tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt tgaagctgat
     1741 gctcgagatc tcggcttgaa cgaattgtta gccaatcgac tggcgagcgg catcgcattc
     1801 ttcgcatccc gcctctggcg gatgcaggaa gatcaacgga tctcggccca gttgacccag
     1861 ggctgtcgcc acaatgtcgc gggagcggat caaccgagca aaggcatgac cgactggacc
     1921 ttccttctga aggctgttct ccttgagcca cctgtccgcc aaggcaaagc gctcacagca
     1981 gtggtcattc tccagataat cgacacgtac caacttgcca tcctgaagaa tggtgcagtg
     2041 tctcggcacc ccatagggaa cctttgccat caactcggca agatgcagcg tcgtgttggc
     2101 atcgtgtccc acgccgagga gaagtacctg cccatcgagt tcatggacac gggcgaccgg
     2161 gcttgcaggc gagtgaggtg gcaggggcaa tggatcagag atgatctgct ctgcctgtgg
     2221 ccccgctgcc gcaaaggcaa atggatgggc gctgcgcttt acatttggca gacgccagaa
     2281 tgtgtcagag acaactccaa ggtccggtgt aacgggcgac gtggcaggat cgaacggctc
     2341 gtcgtccaga cctgaccacg atggcatgac gagcgtccct cccggaccca gtgcagcacg
     2401 cagggcctcg atcagtccaa gtggcccatc ttcgaggggc cggacgctac ggaaggagct
     2461 gtggaccagc agcacaccgc cgggggtaac cccaaggttg agaagctgac cgatcagctc
     2521 ggcttttcgc cattcgtatt gcacgacatt gcactccacc gctgatgaca tcgttgctgc
     2581 tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca
     2641 tagactgtac gaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac
     2701 cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca
     2761 gtttacgaac cgaacaggct tatgtcaact gggttcgtgc cttcatccgt ttccacggtg
     2821 tgcggtccat gagcaaatat tatacgcaag gcgacaaggc tcgagagttc aacagacagc
     2881 cttcgttctt atggccgttc aggaggaagt tcctattccg aagttcctat tctctagaaa
     2941 gtataggaac ttcttgagaa gagaaaagaa aaccgccgat cctgtccacc gcattactgc
     3001 aaggtagtgg acaagaccgg cggtcttaag ttttttggct gaaacgaatg acgaaaaggc
     3061 tgtctgagca aatccaggag gtcgtttaaa ccgtattagc ccggtatttt ggaaatagcg
     3121 gaagcactga cttttgttat caataaaaaa ggccccccgt tagggaggcc ttattgttcg
     3181 tctactcgga agagcgagag acaacagaac ggtcagccac atgaattctt tcagctcagt
     3241 cgataggtag taggcaagag tagtcgcacc tttggtcgaa aaaaaaagcc cgcactgtca
     3301 ggtgcgggct tttttctgtg tttccccaaa agtaaaaacc cgccgaagcg ggtttttacg
     3361 taaaacaggt gaaactgacc agacgagaag gctttggagg tgataatggg gctcaaggac
     3421 ccgggtgggg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     3481 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3541 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     3601 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     3661 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagcta
     3721 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     3781 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     3841 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     3901 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     3961 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     4021 cccaatgcga ccagatgctc cacgcccagt cgcgtaccat cttcatggga gaaaataata
     4081 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4141 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4201 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4261 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     4321 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     4381 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     4441 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     4501 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     4561 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat
     4621 tcgatggtgt caacgtaaat gcatgccgct tccccgactc ggtgccactt tttcgggagg
     4681 cagataggta ggcatggccc ccattttcaa tacaagcaac gcatgagaaa gcccccggaa
     4741 gatcaccttc cgggggcttt tttattgcgc tccggcaatt aaaaaagcgg ctaaccacgc
     4801 cgcttttttt acgtctgcag actagtcagg cagccatcgg aagctgtggt atggctgtgc
     4861 aggtcgtaaa gacgtctcat taatcagata aaatattgac aatgtgagcg gataacaaga
     4921 tactgagcac atcagcagca cacaggaaac agaccatgga attcgagctc ggtacccggg
     4981 gatcctctag agtcgacctg caggcatgca agcttggctg ttttggcgga tgagagaaga
     5041 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     5101 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     5161 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     5221 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     5281 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     5341 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     5401 gccatcctga cggatggcct tttcacagaa tgagcatcat ggcctagtcg acatcggaga
     5461 agagtacggc tcttttaacc gcctcaagaa ccagataagt gaaatctagt tccaaactat
     5521 tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg
     5581 tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat
     5641 tttgtccgcc cacagcgggg catttttctt cctgttatgt ttgggctctt ccgcttcctc
     5701 gctcactgac cctatcagag cttatcggcc agcctcgcag agcaggattc ccgttgagca
     5761 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     5821 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     5881 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     5941 ccccgaagca gggttatgca gcggaaagta taccttaagg aatcccctga taacgcagga
     6001 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcactccct
     6061 gcctctgtca tcacgatact gtgatgccgt ggctaattcc catgtcagcc gttaagtgtt
     6121 cctgtgtcac tcaaaattgc tttgagaggc tctaagggct tctcagtgcg ttacatccct
     6181 ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa agccttatat attctttttt
     6241 ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat taattttatt
     6301 gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag ccatgagagc
     6361 ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta cgttaaacat
     6421 gagagcttag tacgtgaaac atgagagctt agtacgtact atcaacaggt tgaactgctg
     6481 atcttcagat cggccgcaaa aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcaccaata actgcctttg
     6601 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6661 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6721 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6781 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6841 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg
     6901 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6961 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7021 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7081 atcgtggtgt cacgctcgtc gtttggtat
//
