LOCUS       sequence-439068-        7306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 integrative vector with PabstBR promoter expression cassette and
            hygR 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 7306)
  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 7306)
  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: pJMP12089
FEATURES             Location/Qualifiers
     source          1..7306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     174..193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(412..516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     584..602
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(640..662)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     762..781
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1195..1211
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1196..1212
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1240..1334
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1437..1523
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1574..1621
                     /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)."
     promoter        complement(join(2859..2864,2865..2881,2882..2887))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    3004..3051
                     /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(3569..4651)
                     /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     4613..4632
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4652..4729)
                     /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    5057..5113
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5146..5163)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5316..5402
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5494..5521
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     mobile_element  5587..5752
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            5824..5932
                     /note="incP origin of transfer"
     rep_origin      6201..6589
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(6857..7306,1..342,343..411))
                     /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 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
       61 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      121 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      181 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      241 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      301 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      361 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
      421 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      481 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      541 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      601 gccctttcgt ctcgcgcgtt tcggtgacga cggtgaaaac ctctgacaca tgcagctccc
      661 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      721 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      781 actgagagtg caccatgtgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
      841 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      901 ctcttcgcta ttacgccagc tagaggacca gccgcgtaac ctggcaaaat cggttacggt
      961 tgagtaataa atggatgccc tgcgtaagca ggtgtgggcg gacaataaag tcttaaactg
     1021 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact
     1081 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa
     1141 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gggttcgagg
     1201 tcgacggtat cgataagcta gcttaattag ctgagcttgg actcctgttg atagatccag
     1261 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     1321 ttattggtga gaatccaagc tagactgcga tgagtggcag ggcggggcgt aattttttta
     1381 aggcagttat tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa
     1441 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1501 acgctctcct gagtaggaca aatccgccgc taggagcttg cggcccggac gatgagctcg
     1561 aattggggat cttgaagtac ctattccgaa gttcctattc tctagaaagt ataggaactt
     1621 cagagcgctt ttgaagctga tgctcgagat ctcggcttga acgaattgtt attatgcacc
     1681 cggtgcggta tcaggcggac cccacagaaa ctgtgctaat tcttccggat cggtaaaacc
     1741 gctcagatcc agcggtgttt cttcaaacac ttcaaaatca tgcagaaagg taaaggccag
     1801 cagttcacgg gcaaaatctt cggtacgttt ccactgtgca ccatccagca gggctgccag
     1861 aatttcacga tcaccacgaa atgcattcag atgcagctga accaggctat aacggctatc
     1921 accggcataa acatcggtga aatcaacaat accggtaact tcggttgctg ccagatcaac
     1981 aaaaatgttg gtgccatgca gatcaccatg aacaaaacgc ggttcgcgac ctgccagcag
     2041 ggtatcaaca tccggcagcc aatcttccag acggtccagc agacgcggac tcagataacc
     2101 ccaaccacga tgatcttcaa cggttgcggc acgacgttca cgcagcagtt ccggaaaaac
     2161 ttcgctatgc ggtgtcagaa cggtattacc ggtcagcgga acacgatgca gacgacccag
     2221 aacacgtccc agttcacgtg ccagtgccag cagtgcatta cgatcggtgg tgccatccat
     2281 tgcgctacgc caggtggtgc cggtcatacg gctcataacc agatacggcc acggccatgc
     2341 acctgtacca ggacgcagtt caccacgacc taacagacga ggaaccggaa ccggtgcatc
     2401 tgccagaact gcataggctt cgctttcgct tgccaggctt tccggaccac accaatgttc
     2461 accaaacagt ttaataacag gatcaggttc accaaccaga accggattgg tgctttcacc
     2521 cggaacacgc agaacaggcg gaaccggcag acccagttct tccagtgcac gacgtgccag
     2581 cggttcccaa aattcttgat cattgcgcag gcttgcatag ctatcatcac tatcaatacg
     2641 atccagcagc agcaggcttt cttgggtcat cgttgctgct ccataacatc aaacatcgac
     2701 ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtacg aaaaaacagt
     2761 cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc
     2821 tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt
     2881 atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtccatga gcaaatatta
     2941 tacgcaaggc gacaaggctc gagagttcaa cagacagcct tcgttcttat ggccgttcag
     3001 gaggaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt cttgagaaga
     3061 gaaaagaaaa ccgccgatcc tgtccaccgc attactgcaa ggtagtggac aagaccggcg
     3121 gtcttaagtt ttttggctga aacgaatgac gaaaaggctg tctgagcaaa tccaggaggt
     3181 cgtttaaacc gtattagccc ggtattttgg aaatagcgga agcactgact tttgttatca
     3241 ataaaaaagg ccccccgtta gggaggcctt attgttcgtc tactcggaag agcgagagac
     3301 aacagaacgg tcagccacat gaattctttc agctcagtcg ataggtagta ggcaagagta
     3361 gtcgcacctt tggtcgaaaa aaaaagcccg cactgtcagg tgcgggcttt tttctgtgtt
     3421 tccccaaaag taaaaacccg ccgaagcggg tttttacgta aaacaggtga aactgaccag
     3481 acgagaaggc tttggaggtg ataatggggc tcaaggaccc gggtggggtg cctaatgagt
     3541 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3661 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     3721 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     3781 gtttgatggt ggttaacggc gggatataac atgagctatc ttcggtatcg tcgtatccca
     3841 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     3901 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     3961 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     4021 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     4081 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     4141 cgcccagtcg cgtaccatct tcatgggaga aaataatact gttgatgggt gtctggtcag
     4201 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     4261 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     4321 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     4381 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     4441 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     4501 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     4561 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     4621 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     4681 ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtca acgtaaatgc
     4741 atgccgcttc cccgactcgg tgccactttt tcgggaggca gataggtagg catggccccc
     4801 attttcaata caagcaacgc atgagaaagc ccccggaaga tcaccttccg ggggcttttt
     4861 tattgcgctc cggcaattaa aaaagcggct aaccacgccg ctttttttac gtctgcagac
     4921 tagtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaaga cgtctcatta
     4981 atcagataaa atattgacaa tgtgagcgga taacaagata ctgagcacat cagcagcaca
     5041 caggaaacag accatggaat tcgagctcgg tacccgggga tcctctagag tcgacctgca
     5101 ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     5161 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     5221 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     5281 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     5341 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     5401 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     5461 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     5521 tcacagaatg agcatcatgg cctagtcgac atcggagaag agtacggctc ttttaaccgc
     5581 ctcaagaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     5641 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     5701 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     5761 tttttcttcc tgttatgttt gggctcttcc gcttcctcgc tcactgaccc tatcagagct
     5821 tatcggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac
     5881 agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg
     5941 ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc
     6001 gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc
     6061 ggaaagtata ccttaaggaa tcccctgata acgcaggaaa gaacatgtga gcaaaaggcc
     6121 agcaaaaggc caggaaccgt aaaaaggccg cactccctgc ctctgtcatc acgatactgt
     6181 gatgccgtgg ctaattccca tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt
     6241 tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta
     6301 aaccttaaaa gctttaaaag ccttatatat tctttttttt cttataaaac ttaaaacctt
     6361 agaggctatt taagttgctg atttatatta attttattgt tcaaacatga gagcttagta
     6421 cgtgaaacat gagagcttag tacgttagcc atgagagctt agtacgttag ccatgagggt
     6481 ttagttcgtt aaacatgaga gcttagtacg ttaaacatga gagcttagta cgtgaaacat
     6541 gagagcttag tacgtactat caacaggttg aactgctgat cttcagatcg gccgcaaaaa
     6601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6661 ccaatgctta atcagtgagg caccaataac tgcctttgat cttttctacg gggtctgacg
     6721 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6781 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6841 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6901 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6961 gcttaccatc tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag
     7021 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7081 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7141 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7201 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7261 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgt
//
