LOCUS       sequence-439082-        5599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with PabstBR promoter, kanR 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 5599)
  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 5599)
  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: pJMP3653
FEATURES             Location/Qualifiers
     source          1..5599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     375..394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        560..637
                     /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             638..1720
                     /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(657..676)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     misc_feature    1940..1996
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2029..2046)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2199..2285
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2377..2404
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(2480..2485,2486..2502,2503..2508)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2619..3413
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2673..2692)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3283..3302
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     oriT            5036..5145
                     /note="incP origin of transfer"
     rep_origin      join(5528..5599,1..474)
                     /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.
                     "
ORIGIN
        1 gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc
       61 accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc
      121 taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact
      181 caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac
      241 agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa
      301 cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg
      361 agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac
      421 tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc
      481 tttgccgcgg ccctctcact tccctgttaa accggaattc taggcccaaa cggaaggcga
      541 agcggcatgc atttacgttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg
      601 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
      661 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
      721 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
      781 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
      841 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
      901 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
      961 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     1021 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     1081 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     1141 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     1201 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     1261 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     1321 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     1381 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     1441 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     1501 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     1561 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     1621 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1681 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     1741 gttagcgcga attgatctgg tttgacagct tatcatcgac tgcacggtgc accaatgctt
     1801 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa agacgtctca
     1861 ttaatcagat aaaatattga caatgtgagc ggataacaag atactgagca catcagcagc
     1921 acacaggaaa cagaccatgg aattcgagct cggtacccgg ggatcctcta gagtcgacct
     1981 gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     2041 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     2101 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     2161 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     2221 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     2281 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     2341 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     2401 tttttcctga acggccataa gaacgaaggc tgtctgttga actctcgagc cgtggaaacg
     2461 gatgaaggca cgaacccagt tgacataagc ctgttcggtt cgtaaactgt aatgcaagta
     2521 gcgtatgcgc tcaaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     2581 caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2641 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2701 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2761 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     2821 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2881 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2941 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3001 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3061 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3121 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3181 acggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3241 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3301 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3361 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3421 tctggggttc gaaatgactc gagcatcagc ttcaaaagcg ctccgatgta ctgcggtttc
     3481 tggactagtc cgttgacatt atcacagcag gattttaaca ttttgcgttg ttccaaaagt
     3541 tatcaacagc ctagaacgtc ataggaagcg attacagaca ctttatagct atcagcatgg
     3601 gaacataagg gcaggatgaa atatgggtct taaaacgcaa atggtgaggt tttagaggta
     3661 ttttttgaag atgattaagg cggtttgttt ttaaaatttt tggcggctct caggctgctt
     3721 acattttaac cagttcagtg aaaagttctt tttcagcaaa tttctgttta gcaccatagc
     3781 taaaacttgc gtggaacata ttaagaattg accgaaatga cacaatctca attatatttt
     3841 ttttgaaaag ttttctttat caaatatttt aaatcattga tttatatata agtatgcatt
     3901 cattttaata attaatcttt atttaacaat gatttatcta tattcaattg tttaattatt
     3961 cttactaata ttatctctat atcaatattt tttatttaaa aacatatgtt tagtagtgct
     4021 tttgattaaa gtaccagagg gagggagcag agctgaatgg gaaatactca ccctagagcg
     4081 attcttaaaa atcaccctaa agtattccca ttcgatgtac cgtcggtcgg tcgctttcgc
     4141 atcagggatg acatcactgt atcaagctgc cactgttatg attacgattg atagcaccgc
     4201 ctgaacacgc tcataaccgc caattaaatg actactcatt gcgtccgcta ctctgttcag
     4261 ttcccctagt aatagcgttt ttccgatgtg tgctagcgtc actgtacctc atcacccaca
     4321 catggacaga ttgagttaca gacattggct aaatttttgg ggtctatgct tgacaaagcg
     4381 agttaaaacc ttagaattta agacaggtac attaagcccc tgtggtgaaa tcattagcgg
     4441 tcgtcaaacc ttaatgcttt cgattgccga tatgtcagta tcgaagatct gtaccccata
     4501 aattacggta aagccccaag caattgcaag gggcttttat cttttttaac aaaaaaaatt
     4561 tataaatcag gattttataa cactaaataa tccaaagtac atgagtaagt catgaccact
     4621 cctgcgatgt gtgtagcact ctgagtatcc gtattatatc agtgatgtca tatacaacca
     4681 cataatgcgg atgtatcact aactccctag tatctttctg tctgcctgtg cgccccatca
     4741 gtcgattatc aacaagtaaa tctgtctttt cttcaattaa atcatcaatt tcaatagctg
     4801 ctatagggtt gcgttcttca agataatcat agatatttct acgatcttgg cttgcagccc
     4861 ttatgtcctc aggcgcgcac aaaacagcag ggaagcagcg cttttccgct gcataaccct
     4921 gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag
     4981 gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca
     5041 ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc
     5101 acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccggctacc gccggcgtaa
     5161 cagatgaggg caagcggatg gctgatgaaa ccaagccaac caggaagggc agcccaccta
     5221 tcaaggtgta ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg
     5281 gcatgagcct gtcggcctac ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg
     5341 tggactatga gcacgtccgc gagctggccc gcatcaatgg cgacctgggc cgcctgggcg
     5401 gcctgctgaa actctggctc accgacgacc cgcgcacggc gcggttcggt gatgccacga
     5461 tcctcgccct gctggcgaag atcgactcta gctagaaata ttttatctga ttaataagat
     5521 gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg
     5581 ccttgcaggg cggtttttc
//
