LOCUS       sequence-439066-        6985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 integrative vector with PabstBR promoter expression cassette and
            kanR 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 6985)
  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 6985)
  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: pJMP12068
FEATURES             Location/Qualifiers
     source          1..6985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(150..197)
                     /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        join(274..279,280..296,297..302)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             413..1207
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(467..486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1077..1096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(1259..1306)
                     /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)."
     terminator      complement(1357..1443)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1546..1640)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1668..1684)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1669..1685)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2099..2118)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2218..2240
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2278..2296)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2364..2468
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2469..2537,2538..3329)
                     /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"
     primer_bind     complement(2687..2706)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3597..3985
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            4254..4362
                     /note="incP origin of transfer"
     mobile_element  4434..4599
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(4665..4692)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4784..4870)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5023..5040
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     misc_feature    5073..5129
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        5457..5534
                     /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."
     CDS             5535..6617
                     /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     complement(5554..5573)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gggctaatac ggtttaaacg acctcctgga tttgctcaga cagccttttc gtcattcgtt
       61 tcagccaaaa aacttaagac cgccggtctt gtccactacc ttgcagtaat gcggtggaca
      121 ggatcggcgg ttttcttttc tcttctcaag aagttcctat actttctaga gaataggaac
      181 ttcggaatag gaacttcctc ctgaacggcc ataagaacga aggctgtctg ttgaactctc
      241 gagccgtgga aacggatgaa ggcacgaacc cagttgacat aagcctgttc ggttcgtaaa
      301 ctgtaatgca agtagcgtat gcgctcaaaa ctggatggct ttcttgccgc caaggatctg
      361 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
      421 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
      481 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
      541 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
      601 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
      661 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
      721 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
      781 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
      841 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
      901 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
      961 tctcgtcgtg acccacggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     1021 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     1081 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     1141 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     1201 cttctgagcg ggactctggg gttcgaaatg actcgagcat cagcttcaaa agcgctctga
     1261 agttcctata ctttctagag aataggaact tcggaatagg tacttcaaga tccccaattc
     1321 gagctcatcg tccgggccgc aagctcctag cggcggattt gtcctactca ggagagcgtt
     1381 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     1441 ttgatgcctc aagctagaga gtcattaccc caggcgttta agggcaccaa taactgcctt
     1501 aaaaaaatta cgccccgccc tgccactcat cgcagtctag cttggattct caccaataaa
     1561 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     1621 tggatctatc aacaggagtc caagctcagc taattaagct agcttatcga taccgtcgac
     1681 ctcgaacccc acgcccctct ttaatacgac gggcaatttg cacttcagaa aatgaagagt
     1741 ttgctttagc cataacaaaa gtccagtatg ctttttcaca gcataactgg actgatttca
     1801 gtttacaact attctgtcta gtttaagact ttattgtcat agtttagatc tattttgttc
     1861 agtttaagac tttattgtcc gcccacacct gcttacgcag ggcatccatt tattactcaa
     1921 ccgtaaccga ttttgccagg ttacgcggct ggtcctctag ctggcgtaat agcgaagagg
     1981 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     2041 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cacatggtgc actctcagta
     2101 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     2161 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2221 ggagctgcat gtgtcagagg ttttcaccgt cgtcaccgaa acgcgcgaga cgaaagggcc
     2281 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     2341 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2401 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2461 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2521 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2581 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2641 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2701 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2761 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2821 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2881 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2941 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3001 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3061 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3121 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3181 gtgggtcccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3241 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3301 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3361 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3421 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3481 aaaagatcaa aggcagttat tggtgcctca ctgattaagc attggtaact gtcagaccaa
     3541 gtttactcat atatacttta gattgattta aaacttcatt tttaattttt gcggccgatc
     3601 tgaagatcag cagttcaacc tgttgatagt acgtactaag ctctcatgtt tcacgtacta
     3661 agctctcatg tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac
     3721 gtactaagct ctcatggcta acgtactaag ctctcatgtt tcacgtacta agctctcatg
     3781 tttgaacaat aaaattaata taaatcagca acttaaatag cctctaaggt tttaagtttt
     3841 ataagaaaaa aaagaatata taaggctttt aaagctttta aggtttaacg gttgtggaca
     3901 acaagccagg gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttgagtg
     3961 acacaggaac acttaacggc tgacatggga attagccacg gcatcacagt atcgtgatga
     4021 cagaggcagg gagtgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4081 tgttctttcc tgcgttatca ggggattcct taaggtatac tttccgctgc ataaccctgc
     4141 ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg atatacagga
     4201 ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca gccgggcagg
     4261 ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc ttattcgcac
     4321 ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgataagctc tgatagggtc
     4381 agtgagcgag gaagcggaag agcccaaaca taacaggaag aaaaatgccc cgctgtgggc
     4441 ggacaaaata gttgggaact gggaggggtg gaaatggagt ttttaaggat tatttaggga
     4501 agagtgacaa aatagatggg aactgggtgt agcgtcgtaa gctaatacga aaattaaaaa
     4561 tgacaaaata gtttggaact agatttcact tatctggttc ttgaggcggt taaaagagcc
     4621 gtactcttct ccgatgtcga ctaggccatg atgctcattc tgtgaaaagg ccatccgtca
     4681 ggatggcctt ctgcttaatt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     4741 ctccgggccg ttgcttcgca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     4801 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     4861 gttttatttg atgcctggca gttccctact ctcgcatggg gagaccccac actaccatcg
     4921 gcgctacggc gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg
     4981 ccgccaggca aattctgttt tatcagaccg cttctgcgtt ctgatttaat ctgtatcagg
     5041 ctgaaaatct tctctcatcc gccaaaacag ccaagcttgc atgcctgcag gtcgactcta
     5101 gaggatcccc gggtaccgag ctcgaattcc atggtctgtt tcctgtgtgc tgctgatgtg
     5161 ctcagtatct tgttatccgc tcacattgtc aatattttat ctgattaatg agacgtcttt
     5221 acgacctgca cagccatacc acagcttccg atggctgcct gactagtctg cagacgtaaa
     5281 aaaagcggcg tggttagccg cttttttaat tgccggagcg caataaaaaa gcccccggaa
     5341 ggtgatcttc cgggggcttt ctcatgcgtt gcttgtattg aaaatggggg ccatgcctac
     5401 ctatctgcct cccgaaaaag tggcaccgag tcggggaagc ggcatgcatt tacgttgaca
     5461 ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     5521 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     5581 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     5641 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     5701 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     5761 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     5821 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     5881 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     5941 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     6001 tcaacagtat tattttctcc catgaagatg gtacgcgact gggcgtggag catctggtcg
     6061 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     6121 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     6181 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     6241 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     6301 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     6361 ccgaagatag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     6421 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     6481 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     6541 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6601 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccc
     6661 cacccgggtc cttgagcccc attatcacct ccaaagcctt ctcgtctggt cagtttcacc
     6721 tgttttacgt aaaaacccgc ttcggcgggt ttttactttt ggggaaacac agaaaaaagc
     6781 ccgcacctga cagtgcgggc tttttttttc gaccaaaggt gcgactactc ttgcctacta
     6841 cctatcgact gagctgaaag aattcatgtg gctgaccgtt ctgttgtctc tcgctcttcc
     6901 gagtagacga acaataaggc ctccctaacg gggggccttt tttattgata acaaaagtca
     6961 gtgcttccgc tatttccaaa atacc
//
