LOCUS       sequence-439062-        7798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn7 integrative vector with GFP expressed under PabstBR promoter and
            aprR marker on Tn7 transposon..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7798)
  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 7798)
  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: pJMP12049
FEATURES             Location/Qualifiers
     source          1..7798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     617..636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(855..959)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1027..1045
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1083..1105)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1205..1224
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1638..1654
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1639..1655
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1683..1777
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1880..1966
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2017..2064
                     /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(2115..2915)
                     /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(3104..3109,3110..3126,3127..3132))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    3250..3297
                     /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(3815..4897)
                     /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     4859..4878
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4898..4975)
                     /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             join(5300..5302,5303..6013)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             5300..6013
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5330..5358)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5953..5973
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6081..6098)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6251..6337
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6429..6456
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     mobile_element  6522..6687
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            6759..6867
                     /note="incP origin of transfer"
     rep_origin      7136..7524
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(7792..7798,1..785,786..854))
                     /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 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
       61 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
      121 gcaatgatac cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
      181 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
      241 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
      301 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      361 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      421 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      481 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      541 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      601 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      661 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      721 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      781 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      841 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      901 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      961 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     1021 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga cgacggtgaa
     1081 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     1141 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     1201 tatgcggcat cagagcagat tgtactgaga gtgcaccatg tgcggtgtga aataccgcac
     1261 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     1321 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctagagga ccagccgcgt
     1381 aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcaggtgtgg
     1441 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa
     1501 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg
     1561 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt
     1621 aaagaggggc gtggggttcg aggtcgacgg tatcgataag ctagcttaat tagctgagct
     1681 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     1741 gctcggttgc cgccgggcgt tttttattgg tgagaatcca agctagactg cgatgagtgg
     1801 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg
     1861 actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1921 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgctaggagc
     1981 ttgcggcccg gacgatgagc tcgaattggg gatcttgaag tacctattcc gaagttccta
     2041 ttctctagaa agtataggaa cttcagagcg cttttgaagc tgatgctcga gatctcggct
     2101 tgaacgaatt gttagccaat cgactggcga gcggcatcgc attcttcgca tcccgcctct
     2161 ggcggatgca ggaagatcaa cggatctcgg cccagttgac ccagggctgt cgccacaatg
     2221 tcgcgggagc ggatcaaccg agcaaaggca tgaccgactg gaccttcctt ctgaaggctg
     2281 ttctccttga gccacctgtc cgccaaggca aagcgctcac agcagtggtc attctccaga
     2341 taatcgacac gtaccaactt gccatcctga agaatggtgc agtgtctcgg caccccatag
     2401 ggaacctttg ccatcaactc ggcaagatgc agcgtcgtgt tggcatcgtg tcccacgccg
     2461 aggagaagta cctgcccatc gagttcatgg acacgggcga ccgggcttgc aggcgagtga
     2521 ggtggcaggg gcaatggatc agagatgatc tgctctgcct gtggccccgc tgccgcaaag
     2581 gcaaatggat gggcgctgcg ctttacattt ggcagacgcc agaatgtgtc agagacaact
     2641 ccaaggtccg gtgtaacggg cgacgtggca ggatcgaacg gctcgtcgtc cagacctgac
     2701 cacgatggca tgacgagcgt ccctcccgga cccagtgcag cacgcagggc ctcgatcagt
     2761 ccaagtggcc catcttcgag gggccggacg ctacggaagg agctgtggac cagcagcaca
     2821 ccgccggggg taaccccaag gttgagaagc tgaccgatca gctcggcttt tcgccattcg
     2881 tattgcacga cattgcactc caccgctgat gacatcgttg ctgctccata acatcaaaca
     2941 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacgaaaaa
     3001 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3061 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3121 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcggt ccatgagcaa
     3181 atattatacg caaggcgaca aggctcgaga gttcaacaga cagccttcgt tcttatggcc
     3241 gttcaggagg aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcttg
     3301 agaagagaaa agaaaaccgc cgatcctgtc caccgcatta ctgcaaggta gtggacaaga
     3361 ccggcggtct taagtttttt ggctgaaacg aatgacgaaa aggctgtctg agcaaatcca
     3421 ggaggtcgtt taaaccgtat tagcccggta ttttggaaat agcggaagca ctgacttttg
     3481 ttatcaataa aaaaggcccc ccgttaggga ggccttattg ttcgtctact cggaagagcg
     3541 agagacaaca gaacggtcag ccacatgaat tctttcagct cagtcgatag gtagtaggca
     3601 agagtagtcg cacctttggt cgaaaaaaaa agcccgcact gtcaggtgcg ggcttttttc
     3661 tgtgtttccc caaaagtaaa aacccgccga agcgggtttt tacgtaaaac aggtgaaact
     3721 gaccagacga gaaggctttg gaggtgataa tggggctcaa ggacccgggt ggggtgccta
     3781 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3901 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3961 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     4021 aatcctgttt gatggtggtt aacggcggga tataacatga gctatcttcg gtatcgtcgt
     4081 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     4141 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     4201 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     4261 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4321 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4381 gctccacgcc cagtcgcgta ccatcttcat gggagaaaat aatactgttg atgggtgtct
     4441 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4501 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4561 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4621 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4681 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4741 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4801 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4861 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4921 cttccgggcg ctatcatgcc ataccgcgaa aggttttgca ccattcgatg gtgtcaacgt
     4981 aaatgcatgc cgcttccccg actcggtgcc actttttcgg gaggcagata ggtaggcatg
     5041 gcccccattt tcaatacaag caacgcatga gaaagccccc ggaagatcac cttccggggg
     5101 cttttttatt gcgctccggc aattaaaaaa gcggctaacc acgccgcttt ttttacgtct
     5161 gcagactagt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaagacgtc
     5221 tcattaatca gataaaatat tgacaatgtg agcggataac aagatactga gcacatcagc
     5281 agcacacagg aaacagacca tgagcaaagg agaagaactt ttcactggag ttgtcccaat
     5341 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtccgtg gagagggtga
     5401 aggtgatgct acaaacggaa aactcaccct taaatttatt tgcactactg gaaaactacc
     5461 tgttccgtgg ccaacacttg tcactactct gacctatggt gttcaatgct tttcccgtta
     5521 tccggatcac atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     5581 ggaacgcact atatctttca aagatgacgg gacctacaag acgcgtgctg aagtcaagtt
     5641 tgaaggtgat acccttgtta atcgtatcga gttaaagggt attgatttta aagaagatgg
     5701 aaacattctt ggacacaaac tggagtacaa ctttaactca cacaatgtat acatcacggc
     5761 agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaacg ttgaagatgg
     5821 ttccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     5881 tttaccagac aaccattacc tgtcgacaca atctgtcctt tcgaaagatc ccaacgaaaa
     5941 gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac atggcatgga
     6001 tgagctctac aaagatcctc tagagtcgac ctgcaggcat gcaagcttgg ctgttttggc
     6061 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     6121 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     6181 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     6241 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     6301 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     6361 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     6421 aattaagcag aaggccatcc tgacggatgg ccttttcaca gaatgagcat catggcctag
     6481 tcgacatcgg agaagagtac ggctctttta accgcctcaa gaaccagata agtgaaatct
     6541 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     6601 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     6661 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttgggct
     6721 cttccgcttc ctcgctcact gaccctatca gagcttatcg gccagcctcg cagagcagga
     6781 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     6841 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     6901 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     6961 atcgctataa tgaccccgaa gcagggttat gcagcggaaa gtatacctta aggaatcccc
     7021 tgataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7081 ggccgcactc cctgcctctg tcatcacgat actgtgatgc cgtggctaat tcccatgtca
     7141 gccgttaagt gttcctgtgt cactcaaaat tgctttgaga ggctctaagg gcttctcagt
     7201 gcgttacatc cctggcttgt tgtccacaac cgttaaacct taaaagcttt aaaagcctta
     7261 tatattcttt tttttcttat aaaacttaaa accttagagg ctatttaagt tgctgattta
     7321 tattaatttt attgttcaaa catgagagct tagtacgtga aacatgagag cttagtacgt
     7381 tagccatgag agcttagtac gttagccatg agggtttagt tcgttaaaca tgagagctta
     7441 gtacgttaaa catgagagct tagtacgtga aacatgagag cttagtacgt actatcaaca
     7501 ggttgaactg ctgatcttca gatcggccgc aaaaattaaa aatgaagttt taaatcaatc
     7561 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacca
     7621 ataactgcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaa
//
