LOCUS       sequence-439070-        5722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with PabstBR promoter, aprR marker, and replicates
            in A. baumannii and E. coli; empty vector plasmid with MCS for 
            cloning - we recommend NcoI/BamHI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5722)
  AUTHORS   Bacon EE, Tran JS, Nadig N, Peters JM
  TITLE     Modular, inducible, and titratable expression systems for 
            Escherichia coli and Acinetobacter baumannii.
  JOURNAL   Microbiol Spectr. 2024 Nov 5;12(11):e0130624. doi: 
            10.1128/spectrum.01306-24. Epub 2024 Sep 20.
  PUBMED    39302127
REFERENCE   2  (bases 1 to 5722)
  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: "Microbiol 
            Spectr."; date: "2024-11-5"; volume: "12(11):e0130624. doi"; pages: 
            " 10.1128/spectrum.01306-24. Epub 2024 Sep 20"
COMMENT     Sequence Label: pJMP3664
FEATURES             Location/Qualifiers
     source          1..5722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(7..12,13..29,30..35)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             224..1024
                     /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
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKENSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     oriT            2646..2755
                     /note="incP origin of transfer"
     rep_origin      3138..3683
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3584..3603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3769..3846
                     /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."
     CDS             3847..4929
                     /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     complement(3866..3885)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     misc_feature    5149..5205
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5238..5255)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5408..5494
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5586..5613
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 acccagttga cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca
       61 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
      121 atggcttgtt atgactgttt tttcgtacag tctatgcctc gggcatccaa gcagcaagcg
      181 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgtcat cagcggtgga
      241 gtgcaatgtc gtgcaatacg aatggcgaaa agccgagctg atcggtcagc ttctcaacct
      301 tggggttacc cccggcggtg tgctgctggt ccacagctcc ttccgtagcg tccggcccct
      361 cgaagatggg ccacttggac tgatcgaggc cctgcgtgct gcactgggtc cgggagggac
      421 gctcgtcatg ccatcgtggt caggtctgga cgacgagccg ttcgatcctg ccacgtcgcc
      481 cgttacaccg gaccttggag ttgtctctga cacattctgg cgtctgccaa atgtaaagcg
      541 cagcgcccat ccatttgcct ttgcggcagc ggggccacag gcagagcaga tcatctctga
      601 tccattgccc ctgccacctc actcgcctgc aagcccggtc gcccgtgtcc atgaactcga
      661 tgggcaggta cttctcctcg gcgtgggaca cgatgccaac acgacgctgc atcttgccga
      721 gttgatggca aaggttccct atggggtgcc gagacactgc accattcttc aggatggcaa
      781 gttggtacgt gtcgattatc tggagaatga ccactgctgt gagcgctttg ccttggcgga
      841 caggtggctc aaggagaaca gccttcagaa ggaaggtcca gtcggtcatg cctttgctcg
      901 gttgatccgc tcccgcgaca ttgtggcgac agccctgggt caactgggcc gagatccgtt
      961 gatcttcctg catccgccag aggcgggatg cgaagaatgc gatgccgctc gccagtcgat
     1021 tggctaacaa ttcgttcaag ccgagatctc gagcatcagc ttcaaaagcg ctccgatgta
     1081 ctgcggtttc tggactagtc cgttgacatt atcacagcag gattttaaca ttttgcgttg
     1141 ttccaaaagt tatcaacagc ctagaacgtc ataggaagcg attacagaca ctttatagct
     1201 atcagcatgg gaacataagg gcaggatgaa atatgggtct taaaacgcaa atggtgaggt
     1261 tttagaggta ttttttgaag atgattaagg cggtttgttt ttaaaatttt tggcggctct
     1321 caggctgctt acattttaac cagttcagtg aaaagttctt tttcagcaaa tttctgttta
     1381 gcaccatagc taaaacttgc gtggaacata ttaagaattg accgaaatga cacaatctca
     1441 attatatttt ttttgaaaag ttttctttat caaatatttt aaatcattga tttatatata
     1501 agtatgcatt cattttaata attaatcttt atttaacaat gatttatcta tattcaattg
     1561 tttaattatt cttactaata ttatctctat atcaatattt tttatttaaa aacatatgtt
     1621 tagtagtgct tttgattaaa gtaccagagg gagggagcag agctgaatgg gaaatactca
     1681 ccctagagcg attcttaaaa atcaccctaa agtattccca ttcgatgtac cgtcggtcgg
     1741 tcgctttcgc atcagggatg acatcactgt atcaagctgc cactgttatg attacgattg
     1801 atagcaccgc ctgaacacgc tcataaccgc caattaaatg actactcatt gcgtccgcta
     1861 ctctgttcag ttcccctagt aatagcgttt ttccgatgtg tgctagcgtc actgtacctc
     1921 atcacccaca catggacaga ttgagttaca gacattggct aaatttttgg ggtctatgct
     1981 tgacaaagcg agttaaaacc ttagaattta agacaggtac attaagcccc tgtggtgaaa
     2041 tcattagcgg tcgtcaaacc ttaatgcttt cgattgccga tatgtcagta tcgaagatct
     2101 gtaccccata aattacggta aagccccaag caattgcaag gggcttttat cttttttaac
     2161 aaaaaaaatt tataaatcag gattttataa cactaaataa tccaaagtac atgagtaagt
     2221 catgaccact cctgcgatgt gtgtagcact ctgagtatcc gtattatatc agtgatgtca
     2281 tatacaacca cataatgcgg atgtatcact aactccctag tatctttctg tctgcctgtg
     2341 cgccccatca gtcgattatc aacaagtaaa tctgtctttt cttcaattaa atcatcaatt
     2401 tcaatagctg ctatagggtt gcgttcttca agataatcat agatatttct acgatcttgg
     2461 cttgcagccc ttatgtcctc aggcgcgcac aaaacagcag ggaagcagcg cttttccgct
     2521 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
     2581 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
     2641 cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc
     2701 ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccggctacc
     2761 gccggcgtaa cagatgaggg caagcggatg gctgatgaaa ccaagccaac caggaagggc
     2821 agcccaccta tcaaggtgta ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg
     2881 gcggcggccg gcatgagcct gtcggcctac ctgctggccg tcggccaggg ctacaaaatc
     2941 acgggcgtcg tggactatga gcacgtccgc gagctggccc gcatcaatgg cgacctgggc
     3001 cgcctgggcg gcctgctgaa actctggctc accgacgacc cgcgcacggc gcggttcggt
     3061 gatgccacga tcctcgccct gctggcgaag atcgactcta gctagaaata ttttatctga
     3121 ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
     3181 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
     3241 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     3301 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     3361 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3421 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     3481 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     3541 accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc
     3601 tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt
     3661 caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaaa
     3721 ccggaattct aggcccaaac ggaaggcgaa gcggcatgca tttacgttga caccatcgaa
     3781 tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     3841 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     3901 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     3961 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     4021 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     4081 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     4141 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     4201 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     4261 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     4321 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     4381 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     4441 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     4501 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     4561 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     4621 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     4681 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     4741 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     4801 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     4861 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4921 gggcagtgag cgcaacgcaa ttaatgtaag ttagcgcgaa ttgatctggt ttgacagctt
     4981 atcatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt
     5041 atggctgtgc aggtcgtaaa gacgtctcat taatcagata aaatattgac aatgtgagcg
     5101 gataacaaga tactgagcac atcagcagca cacaggaaac agaccatgga attcgagctc
     5161 ggtacccggg gatcctctag agtcgacctg caggcatgca agcttggctg ttttggcgga
     5221 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     5281 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     5341 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     5401 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     5461 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     5521 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     5581 taagcagaag gccatcctga cggatggcct ttttcctgaa cggccataag aacgaaggct
     5641 gtctgttgaa ctctcgagcc ttgtcgcctt gcgtataata tttgctcatg gacgcacacc
     5701 gtggaaacgg atgaaggcac ga
//
