LOCUS       sequence-432290-        5614 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Empty backbone for Tn7-site insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5614)
  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 5614)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  38..203
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            362..470
                     /note="incP origin of transfer"
     rep_origin      complement(702..1290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(782..801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1461..2252,2253..2321))
                     /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     2084..2103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2322..2426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2494..2512
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2550..2572)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2672..2691
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3105..3121
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3106..3122
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3150..3244
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      3347..3433
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    complement(3487..3534)
                     /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(3617..3991)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        4011..4115
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(5368..5415)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    complement(5526..5573)
                     /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)."
ORIGIN
        1 agcttgggcc cggtacctcg cgaaggcctt gcaggccaac cagataagtg aaatctagtt
       61 ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca
      121 tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt
      181 cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt tgggcgctgc
      241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      301 cctcgctcac tgacccgctg cgctcggtcg ttcggctgcg gcgagcggta tcagagctta
      361 tcggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag
      421 tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc
      481 gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga
      541 aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg
      601 aaagtatacc ttaaggaatc ccctgataac gcaggaaaga acatgtgagc aaaaggccag
      661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      721 cctgacgagc atcacaaaaa tcgacgctcg agtcagaggt ggcgaaaccc gacaggacta
      781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1141 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1741 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2461 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     2521 ctcgcgcgtt tcggtgacga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2581 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2641 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2701 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     2761 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     2821 ttacgccagc tagaggacca gccgcgtaac ctggcaaaat cggttacggt tgagtaataa
     2881 atggatgccc tgcgtaagca ggtgtgggcg gacaataaag tcttaaactg aacaaaatag
     2941 atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc
     3001 cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt
     3061 tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gggttcgagg tcgacggtat
     3121 cgataagcta gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc
     3181 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
     3241 gaatccaagc tagactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     3301 tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa
     3361 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     3421 gagtaggaca aatccgccgc taggagcttg cggcccggac gatcgaatta gcttcaaaag
     3481 cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaagatc
     3541 ccctgattcc ctttgtcaac agcaatggat ttatctctga tgttacattg cacaagataa
     3601 aaatatatca tcatgatcag tcctgctcct cggccacgaa gtgcacgcag ttgccggccg
     3661 ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc atggccggcc
     3721 cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac agctcgtcca
     3781 ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc tggaccgcgc
     3841 tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc acgaagtccc
     3901 gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg cgcgcggtga
     3961 gcaccggaac ggcactggtc aacttggcca tgaaagcatg cggaaatgtg cgcggaaccc
     4021 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4081 gataaatgct tcaataatat tgaaaaagga agagtatgag ttgaaaattt tttttttgac
     4141 tcaatatcta gacttgcaag agcttggaac tttgagattg ttctaagatg catacaaatt
     4201 caaaatgctt aaacaaaaat aatttaacaa aggagtaaga ttcatatgac acaagaatcc
     4261 ctgttacttc tcgaccgtat tgattcggat gattcctacg cgagcctgcg gaacgaccag
     4321 gaattctggg agccgctggc ccgccgagcc ctggaggagc tcgggctgcc ggtgccgccg
     4381 gtgctgcggg tgcccggcga gagcaccaac cccgtactgg tcggcgagcc cggcccggtg
     4441 atcaagctgt tcggcgagca ctggtgcggt ccggagagcc tcgcgtcgga gtcggaggcg
     4501 tacgcggtcc tggcggacgc cccggtgccg gtgccccgcc tcctcgtccg cggcgagctg
     4561 cggcccggca ccggagcctg gccgtggccc tacctggtga tgagccggat gaccggcacc
     4621 acctggcggt ccgcgatgga cggcacgacc gaccggaacg cgctgctcgc cctggcccgc
     4681 gaactcggcc gggtgctcgg ccggctgcac agggtgccgc tgaccgggaa caccgtgctc
     4741 accccccatt ccgaggtctt cccggaactg ctgcgggaac gccgcgcggc gaccgtcgag
     4801 gaccaccgcg ggtggggcta cctctcgccc cggctgctgg accgcctgga ggactggctg
     4861 ccggacgtgg acacgctgct ggccggccgc gaaccccggt tcgtccacgg cgacctgcac
     4921 gggaccaaca tcttcgtgga cctggccgcg accgaggtca ccgggatcgt cgacttcacc
     4981 gacgtctatg cgggagaccc ccgctacagc ctggtgcaac tgcatctcaa cgccttccgg
     5041 ggcgaccgcg agatcctggc cgcgctgctc gacggggcgc agtggaagcg gaccgaggac
     5101 ttcgcccgcg aactgctcgc cttcaccttc ctgcacgact tcgaggtgtt cgaggagacc
     5161 ccgctggatc tctccggctt caccgatccg gaggaactgg cgcagttcct ctgggggccg
     5221 ccggacaccg cccccggcgc ctgacaatcg ttcaagccga gatcggcttc ccggccgcgg
     5281 agttgttcgg taaattgtca caacgccgcg gccaattcgc atcgattgtc agaccaagca
     5341 tgctttcatg gtttagttcc tcaccttgtc gtattatact atgccgatat actatgccga
     5401 tgattaattg tcaacacgtg ctgatcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     5461 aagccgaata tctagaatta ttccattgag taagttttta agcacatcag cttcaaaagc
     5521 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggtac ttcaagatcc
     5581 ccaattcgat catgcatgag ctcactagtg gatc
//
