LOCUS       sequence-437320-        6120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for Tn7-site insertion with araC-ParaBAD expression 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6120)
  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 6120)
  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-Para
FEATURES             Location/Qualifiers
     source          1..6120
                     /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(2363..3241)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3268..3552
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3475..3494
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     mobile_element  3637..3802
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3961..4069
                     /note="incP origin of transfer"
     rep_origin      complement(4301..4889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4381..4400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5060..5851,5852..5920))
                     /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     5683..5702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5921..6025)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6093..6111
                     /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 cgaatcccca aattatgaca acttgacggc tacatcattc actttttctt
     2401 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
     2461 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
     2521 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
     2581 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
     2641 cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct
     2701 cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca
     2761 gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc
     2821 cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc
     2881 gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc
     2941 gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt
     3001 gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc
     3061 cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc
     3121 ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg
     3181 ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca
     3241 tacttttcat actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt
     3301 gccgtcactg cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta
     3361 aaagcattct gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct
     3421 ataatcacgg cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca
     3481 tagcattttt atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta
     3541 ctgtttctcc atacccgttt ttttgggcta gactagtgct ctgcaggaat tcctcgagaa
     3601 gcttgggccc ggtacctcgc gaaggccttg caggccaacc agataagtga aatctagttc
     3661 caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat
     3721 ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc
     3781 ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt gggcgctgca
     3841 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3901 ctcgctcact gacccgctgc gctcggtcgt tcggctgcgg cgagcggtat cagagcttat
     3961 cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt
     4021 gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg
     4081 ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa
     4141 aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga
     4201 aagtatacct taaggaatcc cctgataacg caggaaagaa catgtgagca aaaggccagc
     4261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4321 ctgacgagca tcacaaaaat cgacgctcga gtcagaggtg gcgaaacccg acaggactat
     4381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4741 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4981 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5041 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5101 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5161 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5221 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5281 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5341 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5401 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5461 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5521 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5581 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5641 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5701 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5761 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5821 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5881 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5941 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6001 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     6061 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
