LOCUS       sequence-437321-        6437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for Tn7-site insertion with lacIq-Ptac expression 
            system and C-terminal VSVG fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6437)
  AUTHORS   Intorcia V, Sava RL, Schroeder GP, Gebhardt MJ
  TITLE     A series of vectors for inducible gene expression in 
            multidrug-resistant Acinetobacter baumannii.
  JOURNAL   Appl Environ Microbiol. 2024 Sep 18;90(9):e0047424. doi: 
            10.1128/aem.00474-24. Epub 2024 Aug 20.
  PUBMED    39162403
REFERENCE   2  (bases 1 to 6437)
  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: "Appl 
            Environ Microbiol."; date: "2024-09-18"; volume: "90(9):e0047424. 
            doi"; pages: " 10.1128/aem.00474-24. Epub 2024 Aug 20"
COMMENT     Sequence Label: pUC18T-mini-Tn7T-hph-Ptac-VSVG
FEATURES             Location/Qualifiers
     source          1..6437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     584..600
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      826..912
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    958..1005
                     /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)."
     protein_bind    2266..2313
                     /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(2390..3472)
                     /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     3434..3453
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3473..3550)
                     /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."
     promoter        join(3780..3785,3786..3801,3802..3808)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3816..3832
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     primer_bind     3821..3843
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             3874..3906
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     mobile_element  3954..4119
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            4278..4386
                     /note="incP origin of transfer"
     rep_origin      complement(4618..5206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4698..4717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5377..6168,6169..6237))
                     /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     6000..6019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6238..6342)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6410..6428
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgacgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
      361 tggatgccct gcgtaagcag gtgtgggcgg acaataaagt cttaaactga acaaaataga
      421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
      481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
      541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacggtatc
      601 gataagctag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      721 aatccaagct agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
      781 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      901 agtaggacaa atccgccgct aggagcttgc ggcccggacg atcgaattgg ggatcttgaa
      961 gtacctattc cgaagttcct attctctaga aagtatagga acttcagcta gagggggcgt
     1021 caggcgccgg gggcggtgtc cggcggcccc cagaggaact gcgccagttc ctccggatcg
     1081 gtgaagccgg agagatccag cggggtctcc tcgaacacct cgaagtcgtg caggaaggtg
     1141 aaggcgagca gttcgcgggc gaagtcctcg gtccgcttcc actgcgcccc gtcgagcagc
     1201 gcggccagga tctcgcggtc gccccggaag gcgttgagat gcagttgcac caggctgtag
     1261 cgggagtctc ccgcatagac gtcggtgaag tcgacgatcc cggtgacctc ggtcgcggcc
     1321 aggtccacga agatgttggt cccgtgcagg tcgccgtgga cgaaccgggg ttcgcggccg
     1381 gccagcagcg tgtccacgtc cggcagccag tcctccaggc ggtccagcag ccggggcgag
     1441 aggtagcccc acccgcggtg gtcctcgacg gtcgccgcgc ggcgttcccg cagcagttcc
     1501 gggaagacct cggaatgggg ggtgagcacg gtgttcccgg tcagcggcac cctgtgcagc
     1561 cggccgagca cccggccgag ttcgcgggcc agggcgagca gcgcgttccg gtcggtcgtg
     1621 ccgtccatcg cggaccgcca ggtggtgccg gtcatccggc tcatcaccag gtagggccac
     1681 ggccaggctc cggtgccggg ccgcagctcg ccgcggccga ggaggcgggg caccggcacc
     1741 ggggcgtccg ccaggaccgc gtacgcctcc gactccgacg cgaggctctc cggaccgcac
     1801 cagtgctcgc cgaacagctt gatcaccggg ccgggctcgc cgaccagtac ggggttggtg
     1861 ctctcgccgg gcacccgcag caccggcggc accggcagcc cgagctcctc cagggctcgg
     1921 cgggccagcg gctcccagaa ttcctggtcg ttccgcaggc tcgcgtagga atcatccgaa
     1981 tcaatacggt cgagaagtaa cagggattct tgtgtcacag cggacctcta ttcacagggt
     2041 acgggccggc ttaattccgc acggccggtc gcgacacggc ctgtccgcac cgcggatcag
     2101 gcgttgacga tgacgggctg gtcggccacg tcggggagtc aggcaactat ggatgaacga
     2161 aatagacaga tcgctgatca gcggcccgga aacgtcctcg aaacgacgca tgtgttcctc
     2221 ctggttggta caggtggttg ggggtgctcg gctgtcgctg gtgttgaagt tcctattccg
     2281 aagttcctat tctctagaaa gtataggaac ttcagagcgc ttttgaagct aattcgatca
     2341 tgcatgagct ccttgcaatt cgcgctaact tacattaatt gcgttgcgct cactgcccgc
     2401 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2461 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     2521 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     2581 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     2641 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
     2701 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     2761 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     2821 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     2881 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     2941 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     3001 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     3061 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     3121 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     3181 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     3241 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     3301 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     3361 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     3421 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     3481 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt
     3541 cgatggtgtc aacgtaaatg ccgcttcgcc ttcgcgcgcg aattgcaagc tgatccgggc
     3601 ttatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta
     3661 tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct
     3721 ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga aatgagctgt
     3781 tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac aatttcacac
     3841 aggaaacaga ctagtgctct gcaggcggcc gcttacacag atattgaaat gaaccgttta
     3901 ggtaagtaat aatgaaagct tgggcccggt acctcgcgaa ggccttgcag gccaaccaga
     3961 taagtgaaat ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg
     4021 ctacacccag ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt
     4081 tccacccctc ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt
     4141 tatgtttggg cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4201 gggcgctctt ccgcttcctc gctcactgac ccgctgcgct cggtcgttcg gctgcggcga
     4261 gcggtatcag agcttatcgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
     4321 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     4381 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
     4441 cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag
     4501 cagggttatg cagcggaaag tataccttaa ggaatcccct gataacgcag gaaagaacat
     4561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcgagtc agaggtggcg
     4681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4801 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5041 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5461 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5761 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6121 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6181 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6241 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6301 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6361 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6421 cacgaggccc tttcgtc
//
