LOCUS       sequence-432291-        9180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA expression plasmid for CRISPR-intereference.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9180)
  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 9180)
  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: pWH1266-Apra-sgRNA
FEATURES             Location/Qualifiers
     source          1..9180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(77..665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(157..176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1167..1967)
                     /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
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     promoter        join(2258..2263,2264..2280,2281..2286)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2634..2668
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        2689..2764
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             2785..2814
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(8823..8845)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    8931..9071
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     8986..9005
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(9086..9103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
       61 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      481 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
      541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
      721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
      781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
      841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1141 ttgccattgc tgcagtcatg agctcagcca atcgactggc gagcggcatc gcattcttcg
     1201 catcccgcct ctggcggatg caggaagatc aacggatctc ggcccagttg acccagggct
     1261 gtcgccacaa tgtcgcggga gcggatcaac cgagcaaagg catgaccgac tggaccttcc
     1321 ttctgaaggc tcttctcctt gagccacctg tccgccaagg caaagcgctc acagcagtgg
     1381 tcattctcga gataatcgac gcgtaccaac ttgccatcct gaagaatggt gcagtgtctc
     1441 ggcaccccat agggaacctt tgccatcaac tcggcaagat gcagcgtcgt gttggcatcg
     1501 tgtcccacgc cgaggagaag tacctgccca tcgagttcat ggacacgggc gaccgggctt
     1561 gcaggcgagt gaggtggcag gggcaatgga tcagagatga tctgctctgc ctgtggcccc
     1621 gctgccgcaa aggcaaatgg atgggcgctg cgctttacat ttggcaggcg ccagaatgtg
     1681 tcagagacaa ctccaaggtc cggtgtaacg ggcgacgtgg caggatcgaa cggctcgtcg
     1741 tccagacctg accacgaggg catgacgagc gtccctcccg gacccagcgc agcacgcagg
     1801 gcctcgatca gtccaagtgg cccatcttcg aggggccgga cgctacggaa ggagctgtgg
     1861 accagcagca caccgccggg ggtaacccca aggttgagaa gctgaccgat gagctcggct
     1921 tttcgccatt cgtattgcac gacattgcac tccaccgctg atgacatcag tcgatcatag
     1981 cacgatcaac ggcactgttg caaatagtag gtggtgataa acttatcatc cccttttgct
     2041 gatggagctg cacatgaacc cattcaaagg ccggcatttt cagcgtgaca tcattctgtg
     2101 ggccgtacgc tggtactgca aatacggcat cagttaccgt gagccggatc agtgagggtt
     2161 tgcaactgcg ggtcaaggat ctggatttcg atcacggcac gatcatcgtg cgggagggca
     2221 agggctccaa ggatcgggcc ttgatgaatt ctcatgtttg acagcttatc atcgataagc
     2281 tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat
     2341 ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta ggcataggct
     2401 tggttatgcc ggtactgccg ggcctcttgc gggatatcgt ccattccgac agcatcgcca
     2461 gtcactatgg cgtgctgcta gcgctatatg cgttgatgca atttctatgc gcacccgttc
     2521 tcggagcact gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg ctacttggag
     2581 ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggatccga tctttgacag
     2641 ctagctcagt cctaggtata atactagtaa ctttcagttt agcggtctgt tttagagcta
     2701 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     2761 gtgctttttt tgaagcttgg gcccgaacaa aaactcatct cagaagagga tctgaatagc
     2821 gccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     2881 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     2941 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     3001 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     3061 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
     3121 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
     3181 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     3241 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt
     3301 cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg
     3361 ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
     3421 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
     3481 caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa
     3541 cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc
     3601 gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     3661 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     3721 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     3781 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
     3841 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     3901 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
     3961 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
     4021 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
     4081 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     4141 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     4201 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgctgctgtt gctgctgttc
     4261 aagtgtcaat tgctctactc tgttgctcgt ctcaatgata gacatatcaa gcctttcagc
     4321 ttttttattg cgctttttga tttcatcatt acgcttgacg atctccagtt cctcccccct
     4381 gcgcttggcg tgggtggctt caagtccctc cttgaatgtc ggctcgatgt ctagtccacg
     4441 atccttatga ctgcggtgat caactctcac ctcaagccct gctcgctcta ggtggacgtt
     4501 cgtcagatcc gccacttttt ccctgatttt tttcagcgtt gagttctgat ctagctccct
     4561 gactttcttc cctagccctt gcggtgtaag gcttcttgtt gtcattagta tgtgggcgtg
     4621 atggtttctt tcatcacttc ccccatgcac atgaggggca tggatcgcta cgtccaccgc
     4681 cactccatag gctttaacta aggactggca caactcggta acaagtgcct gacgctgtgt
     4741 cttatccagt tcatgcggca atgctatctc tacttccttt gctaatcttg cctcctgttt
     4801 gatgtcaccg ttctttttta gttcggattg ctctacccga ttccataggg tttgacgatc
     4861 taacatgtca ggacttgccc cagttggggc aaaaatttgg gtgtattcaa tgcctgtttt
     4921 tttggtgtag tcctgctctt ttccgtacgt atcacagtac aatttttcgc ctgctcggta
     4981 tgctgcacat gccacgattg agcgaccatc tgacctcgaa atgttctgca tttcacaatg
     5041 gtaaattgcc attttcatca cctgtttatt aacagccgcc ccaagttttg ggcggttggg
     5101 gtgtcggggt ttccctgacc gtacttgcaa aatttggcgt agcaaaaatt ttgcgtaagt
     5161 gcgcccttcg ggaactctgc gaggctaaaa gcaaaagcaa gagcaataag gctttgactt
     5221 tgttttttga ttttgctcgc tgcgctcgga tcttgatgta agtcgagttt ttgaagtaga
     5281 acacttttca cattgatgat ggttcatact tcggaaaata ggttgagaca caggcattaa
     5341 aaatggtcaa aaaaacagct atggaattgg gtcaaatgct cgatgaagaa aaggaaaaat
     5401 tggaacgtca ggaaaaggca agaaaggatc tagctgacgc tgtggtgaaa ggtcgagagc
     5461 aaaaacaggc aaggtcagac gatgcaaaac gcaagatcct gatcggggct tacttactga
     5521 agaaacatga taatgacatc aagaaacttg tggaacaaaa ccccgatttt attgggtaca
     5581 ttagagaaaa tgataagcac ttattttcat aggcattagt agttataaaa aaccctcata
     5641 aatcgagggt ttctttttta tctgcattac tttttgaatc aatgtacttt tttagcttgc
     5701 tatttttcac tgcatacagc cgcttatact ctttctcaat caatgcggtc aaaatggctt
     5761 tttggctctc ccctgctgcg cccaccatgt ccgcaagcat atccgacaca cccttatcta
     5821 taaacacctc caaacgttga ctatcaaggc ttttgcgttt ttcacgatat gctttttgtc
     5881 gttcggcatt ggtttttggc ggctctttta gcatgtcctg tgttttctga tcgttcgggt
     5941 ctttcatgtc tttaactccc atatagtata tcatagttac aggataacat tgtttttagt
     6001 tacaggataa ctttttataa aatttatttg taaaaattta tgctatcctg caactatttt
     6061 taaattacag gataactatt taacgttacc ctgtaactat agttatcctg taacgctaaa
     6121 ttaattaccc tgtaacatat attttgttat cctgtaacaa ctaaatattg aaataatact
     6181 accaaagtta ccctgtaacg aaaataacta ggttaaaaaa tgatcggatt ttaacatttt
     6241 gcgttgttcc aaaagttatc aacagcctag aacgtcatag gaagcgatta cagacacttt
     6301 atagctatca gcatgggaac ataagggcag gatgaaatat gggtcttaaa acgcaaatgg
     6361 tgaggtttta gaggtatttt ttgaagatga ttaaggcggt ttgtttttaa aatttttggc
     6421 ggctctcagg ctgcttacat tttaaccagt tcagtgaaaa gttctttttc agcaaatttc
     6481 tgtttagcac catagctaaa acttgcgtgg aacatattaa gaattgaccg aaatgacaca
     6541 atctcaatta tatttttttt gaaaagtttt ctttatcaaa tattttaaat cattgattta
     6601 tatataagta tacattcatt ttaataatta atctttattt aacaatgatt tatctatatt
     6661 caattgttta attattctta ctaatattat ctctatatca atatttttta tttaaaaaca
     6721 tatgtttagt agtgcttttg attaaagtac cagagggagg gagcagagct gaatgggaaa
     6781 tactcaccct agagcgattc ttaaaaatca ccctaaagta ttcccattcg atgtaccgtc
     6841 ggtcggtcgc tttcgcatca gggatgacat cactgtatca agctgccact gttatgatta
     6901 cgattgatag caccgcctga acacgctcat aaccgccaat taaatgacta ctcattgcgt
     6961 ccgctactct gttcagttcc cctagtaata gcgtttttcc gatgtgtgct agcgtcactg
     7021 tacctcatca cccacacatg gacagattga gttacagaca ttggctaaat ttttggggtc
     7081 tatgcttgac aaagcgagtt aaaaccttag aatttaagac aggtacatta agcccctgtg
     7141 gtgaaatcat tagcggtcgt caaaccttaa tgctttcgat tgccgatatg tcagtatcga
     7201 agatctgtac cccataaatt acggtaaagc cccaagcaat tgcaaggggc ttttatcttt
     7261 tttaacaaaa aaaatttata aatcaggatt ttataacact aaataatcca aagtacatga
     7321 gtaagtcatg accactcctg cgatgtgtgt agcactctga gtatccgtat tatatcagtg
     7381 atgtcatata caaccacata atgcggatgt atcactaact ccctagtatc tttctgtctg
     7441 cctgtgcgcc ccatcagtcg attatcaaca agtaaatctg tcttttcttc aattaaatca
     7501 tcaatttcaa tagctgctat agggttgcgt tcttcaagat aatcatagat atttctacga
     7561 tcttggcgag ctgtttcagt ccactcaatc ttcattcttc gctgccagtt tcgccaatgt
     7621 tgcagcacgt ctagcggcaa actctgcctt tacgtcctca tgtggaatca acttgcctgc
     7681 attggcttca tttaagccga tctgtacctg ctgcttgaac catttgtcat agtctgcggc
     7741 ttcctgttgc ttcttaacaa agtcacgcat gaagtcacgg attagctgcg cccccgaccg
     7801 atcaacagtc ttagcaaggc ttgaaaattc ttgctttagg tcatggtcta cccgaaaggt
     7861 aaaagtcgct tctgtcatgt ttttcactcc aatgtgttac atttgaatgc tattgtaaca
     7921 cattaaaaag ttaggtcatt gtcttttttc ggcttaggct gttcttggtc ataacgtggt
     7981 ttaaatgatg gctgtcgctc attctcaagt aaaattttcc gtttattaac agcctgttgc
     8041 atctgctcat agtaaggcat caactgcggt aaaatatcag caagtgcctg tctgtcttta
     8101 tccaggatac ggctcaattg cccgattttt ttcaagtcac gctcataggc tattctgtcg
     8161 ctgatctgct cgtatttttg ctttgaacac cttatgtact cttgcatatc gttacttgcc
     8221 catgcctgcg cctgaagacg tgtaaacatg gaaatgtctg aagcaacatg gtcgctgata
     8281 tggtcatact cagcatcttt gacatagccg tattttgata aatatctttc atagtcctgc
     8341 cgtttctttt tgtcatcttc ctgtttttgt ttagtttctc tttctatctg ctgttgtttt
     8401 cgttgctgtt cctgttgctc ctcttttatc tctctgtgat aaatggaaag gctagattga
     8461 tcattatccc aacgtgtgag catgtcctca aagccgtaat aatcagcact aaaccaacct
     8521 gttttcactt tcactggtgg cggcagtggc tctccgatct cccttagcag tgctgtagct
     8581 gctcgtccct ctttgatcac tttatcaagc tgcaagcggt cgatattgtc ttgcaaagcc
     8641 ttgtaaaatc tatgctctgc tcgcttctca tccgttttta cttcaatcaa atacacatct
     8701 ataagcgttt ggcggtgttg ttggtggtcg tatagtgcct gttgtgcggt ggcatagact
     8761 gcgggggctt ggtctacggc atcctccagc tgcctcgcgc gtttcggtga tgacggtgaa
     8821 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     8881 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     8941 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     9001 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     9061 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     9121 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
//
