LOCUS       sequence-432230-        8171 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Empty backbone for lacZ fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8171)
  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 8171)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-lacZ
FEATURES             Location/Qualifiers
     source          1..8171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    221..268
                     /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)."
     primer_bind     486..502
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     486..502
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             498..3569
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="MTMIKLAPVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTD
                     RPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVT
                     YPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQ
                     DSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQI
                     SDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDE
                     RGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENG
                     LLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWY
                     TLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSL
                     GNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPK
                     WSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLI
                     KYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQT
                     IEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQL
                     WLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGN
                     KRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKA
                     AGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVE
                     VASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYIPYV
                     FPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGF
                     HMGIGGDDSWSPSVSAELQLSAGRYHYQLVWCQK"
     primer_bind     complement(524..546)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     mobile_element  3643..3808
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3967..4075
                     /note="incP origin of transfer"
     rep_origin      complement(4307..4895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4387..4406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5066..5857,5858..5926))
                     /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     5689..5708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5927..6031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6099..6117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6155..6177)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6277..6296
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6710..6726
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6711..6727
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6755..6849
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      6952..7038
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    7084..7131
                     /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 ccggcttaat tccgcacggc cggtcgcgac acggcctgtc cgcaccgcgg atcaggcgtt
       61 gacgatgacg ggctggtcgg ccacgtcggg gagtcaggca actatggatg aacgaaatag
      121 acagatcgct gatcagcggc ccggaaacgt cctcgaaacg acgcatgtgt tcctcctggt
      181 tggtacaggt ggttgggggt gctcggctgt cgctggtgtt gaagttccta ttccgaagtt
      241 cctattctct agaaagtata ggaacttcag agcgcttttg aagctaattc gatcatgcat
      301 gagctcgaat tcccggggat ccgcatgcct gcagtaacta actagcgatc ccgactcact
      361 atagagggac aaactcaagg tcattcgcaa gagtggcctt tatgattgac cttcttccgg
      421 ttaatacgac cgggatcgag atcctaggta ggtaggggcg cggcatttta actttcttta
      481 tcacacagga aacagctatg accatgatta agcttgctcc cgtcgtttta caacgtcgtg
      541 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      601 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      661 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
      721 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
      781 acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc
      841 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
      901 aggaaggcca gacgcgaatt atttttgatg gcgttaactc ggcgtttcat ctgtggtgca
      961 acgggcgctg ggtcggttac ggccaggaca gtcgtttgcc gtctgaattt gacctgagcg
     1021 catttttacg cgccggagaa aaccgcctcg cggtgatggt gctgcgctgg agtgacggca
     1081 gttatctgga agatcaggat atgtggcgga tgagcggcat tttccgtgac gtctcgttgc
     1141 tgcataaacc gactacacaa atcagcgatt tccatgttgc cactcgcttt aatgatgatt
     1201 tcagccgcgc tgtactggag gctgaagttc agatgtgcgg cgagttgcgt gactacctac
     1261 gggtaacagt ttctttatgg cagggtgaaa cgcaggtcgc cagcggcacc gcgcctttcg
     1321 gcggtgaaat tatcgatgag cgtggtggtt atgccgatcg cgtcacacta cgtctgaacg
     1381 tcgaaaaccc gaaactgtgg agcgccgaaa tcccgaatct ctatcgtgcg gtggttgaac
     1441 tgcacaccgc cgacggcacg ctgattgaag cagaagcctg cgatgtcggt ttccgcgagg
     1501 tgcggattga aaatggtctg ctgctgctga acggcaagcc gttgctgatt cgaggcgtta
     1561 accgtcacga gcatcatcct ctgcatggtc aggtcatgga tgagcagacg atggtgcagg
     1621 atatcctgct gatgaagcag aacaacttta acgccgtgcg ctgttcgcat tatccgaacc
     1681 atccgctgtg gtacacgctg tgcgaccgct acggcctgta tgtggtggat gaagccaata
     1741 ttgaaaccca cggcatggtg ccaatgaatc gtctgaccga tgatccgcgc tggctaccgg
     1801 cgatgagcga acgcgtaacg cgaatggtgc agcgcgatcg taatcacccg agtgtgatca
     1861 tctggtcgct ggggaatgaa tcaggccacg gcgctaatca cgacgcgctg tatcgctgga
     1921 tcaaatctgt cgatccttcc cgcccggtgc agtatgaagg cggcggagcc gacaccacgg
     1981 ccaccgatat tatttgcccg atgtacgcgc gcgtggatga agaccagccc ttcccggctg
     2041 tgccgaaatg gtccatcaaa aaatggcttt cgctacctgg agagacgcgc ccgctgatcc
     2101 tttgcgaata cgcccacgcg atgggtaaca gtcttggcgg tttcgctaaa tactggcagg
     2161 cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc
     2221 tgattaaata tgatgaaaac ggcaacccgt ggtcggctta cggcggtgat tttggcgata
     2281 cgccgaacga tcgccagttc tgtatgaacg gtctggtctt tgccgaccgc acgccgcatc
     2341 cagcgctgac ggaagcaaaa caccagcagc agtttttcca gttccgttta tccgggcaaa
     2401 ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga taacgagctc ctgcactgga
     2461 tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt gcctctggat gtcgctccac
     2521 aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc ggagagcgcc gggcaactct
     2581 ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg gtcagaagcc gggcacatca
     2641 gcgcctggca gcagtggcgt ctggcggaaa acctcagtgt gacgctcccc gccgcgtccc
     2701 acgccatccc gcatctgacc accagcgaaa tggatttttg catcgagctg ggtaataagc
     2761 gttggcaatt taaccgccag tcaggctttc tttcacagat gtggattggc gataaaaaac
     2821 aactgctgac gccgctgcgc gatcagttca cccgtgcacc gctggataac gacattggcg
     2881 taagtgaagc gacccgcatt gaccctaacg cctgggtcga acgctggaag gcggcgggcc
     2941 attaccaggc cgaagcagcg ttgttgcagt gcacggcaga tacacttgct gatgcggtgc
     3001 tgattacgac cgctcacgcg tggcagcatc aggggaaaac cttatttatc agccggaaaa
     3061 cctaccggat tgatggtagt ggtcaaatgg cgattaccgt tgatgttgaa gtggcgagcg
     3121 atacaccgca tccggcgcgg attggcctga actgccagct ggcgcaggta gcagagcggg
     3181 taaactggct cggattaggg ccgcaagaaa actatcccga ccgccttact gccgcctgtt
     3241 ttgaccgctg ggatctgcca ttgtcagaca tgtatatccc gtacgtcttc ccgagcgaaa
     3301 acggtctgcg ctgcgggacg cgcgaattga attatggccc acaccagtgg cgcggcgact
     3361 tccagttcaa catcagccgc tacagtcaac agcaactgat ggaaaccagc catcgccatc
     3421 tgctgcacgc ggaagaaggc acatggctga atatcgacgg tttccatatg gggattggtg
     3481 gcgacgactc ctggagcccg tcagtatcgg cggaattaca gctgagcgcc ggtcgctacc
     3541 attaccagtt ggtctggtgt caaaaataat aataaccggg caggccatgt ctgcccgtat
     3601 ttcgcgtaag gaaatccatt atgtactatt tccttgcagg ccaaccagat aagtgaaatc
     3661 tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt
     3721 tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc
     3781 cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttgggc
     3841 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3901 cgcttcctcg ctcactgacc cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcaga
     3961 gcttatcggc cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg
     4021 gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg
     4081 acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg
     4141 tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc
     4201 agcggaaagt ataccttaag gaatcccctg ataacgcagg aaagaacatg tgagcaaaag
     4261 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4321 gcccccctga cgagcatcac aaaaatcgac gctcgagtca gaggtggcga aacccgacag
     4381 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4441 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4501 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4561 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4621 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4681 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4741 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4801 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5941 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6001 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6061 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6121 ttcgtctcgc gcgtttcggt gacgacggtg aaaacctctg acacatgcag ctcccggaga
     6181 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6241 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6301 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6361 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     6421 cgctattacg ccagctagag gaccagccgc gtaacctggc aaaatcggtt acggttgagt
     6481 aataaatgga tgccctgcgt aagcaggtgt gggcggacaa taaagtctta aactgaacaa
     6541 aatagatcta aactatgaca ataaagtctt aaactagaca gaatagttgt aaactgaaat
     6601 cagtccagtt atgctgtgaa aaagcatact ggacttttgt tatggctaaa gcaaactctt
     6661 cattttctga agtgcaaatt gcccgtcgta ttaaagaggg gcgtggggtt cgaggtcgac
     6721 ggtatcgata agctagctta attagctgag cttggactcc tgttgataga tccagtaatg
     6781 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
     6841 ggtgagaatc caagctagac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca
     6901 gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca tcaaataaaa
     6961 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     7021 ctcctgagta ggacaaatcc gccgctagga gcttgcggcc cggacgatcg aattggggat
     7081 cttgaagtac ctattccgaa gttcctattc tctagaaagt ataggaactt cagctagagg
     7141 gggcgtcagg cgccgggggc ggtgtccggc ggcccccaga ggaactgcgc cagttcctcc
     7201 ggatcggtga agccggagag atccagcggg gtctcctcga acacctcgaa gtcgtgcagg
     7261 aaggtgaagg cgagcagttc gcgggcgaag tcctcggtcc gcttccactg cgccccgtcg
     7321 agcagcgcgg ccaggatctc gcggtcgccc cggaaggcgt tgagatgcag ttgcaccagg
     7381 ctgtagcggg agtctcccgc atagacgtcg gtgaagtcga cgatcccggt gacctcggtc
     7441 gcggccaggt ccacgaagat gttggtcccg tgcaggtcgc cgtggacgaa ccggggttcg
     7501 cggccggcca gcagcgtgtc cacgtccggc agccagtcct ccaggcggtc cagcagccgg
     7561 ggcgagaggt agccccaccc gcggtggtcc tcgacggtcg ccgcgcggcg ttcccgcagc
     7621 agttccggga agacctcgga atggggggtg agcacggtgt tcccggtcag cggcaccctg
     7681 tgcagccggc cgagcacccg gccgagttcg cgggccaggg cgagcagcgc gttccggtcg
     7741 gtcgtgccgt ccatcgcgga ccgccaggtg gtgccggtca tccggctcat caccaggtag
     7801 ggccacggcc aggctccggt gccgggccgc agctcgccgc ggccgaggag gcggggcacc
     7861 ggcaccgggg cgtccgccag gaccgcgtac gcctccgact ccgacgcgag gctctccgga
     7921 ccgcaccagt gctcgccgaa cagcttgatc accgggccgg gctcgccgac cagtacgggg
     7981 ttggtgctct cgccgggcac ccgcagcacc ggcggcaccg gcagcccgag ctcctccagg
     8041 gctcggcggg ccagcggctc ccagaattcc tggtcgttcc gcaggctcgc gtaggaatca
     8101 tccgaatcaa tacggtcgag aagtaacagg gattcttgtg tcacagcgga cctctattca
     8161 cagggtacgg g
//
