LOCUS       sequence-439080-        6609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with GFP expressed under PabstBR promoter, hygR 
            marker, and replicates in A. baumannii and E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6609)
  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 6609)
  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: pJMP3667
FEATURES             Location/Qualifiers
     source          1..6609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            134..243
                     /note="incP origin of transfer"
     promoter        complement(join(3052..3057,3058..3074,3075..3080))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     terminator      complement(3196..3223)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3315..3401)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3554..3571
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(3639..4349,4350..4352))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(3639..4352)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3679..3699)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     4294..4322
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(4569..5651)
                     /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     5613..5632
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5652..5729)
                     /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."
     rep_origin      complement(5815..6360)
                     /direction=LEFT
                     /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     complement(5895..5914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccgccgccg ccttttcctc aatcgctctt cgttcgtctg gaaggcagta caccttgata
       61 ggtgggctgc ccttcctggt tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt
      121 acgccggcgg tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc
      181 gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg
      241 cccggctgac gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct
      301 gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag
      361 ggttatgcag cggaaaagcg ctgcttccct gctgttttgt gcgcgcctga ggacataagg
      421 gctgcaagcc aagatcgtag aaatatctat gattatcttg aagaacgcaa ccctatagca
      481 gctattgaaa ttgatgattt aattgaagaa aagacagatt tacttgttga taatcgactg
      541 atggggcgca caggcagaca gaaagatact agggagttag tgatacatcc gcattatgtg
      601 gttgtatatg acatcactga tataatacgg atactcagag tgctacacac atcgcaggag
      661 tggtcatgac ttactcatgt actttggatt atttagtgtt ataaaatcct gatttataaa
      721 ttttttttgt taaaaaagat aaaagcccct tgcaattgct tggggcttta ccgtaattta
      781 tggggtacag atcttcgata ctgacatatc ggcaatcgaa agcattaagg tttgacgacc
      841 gctaatgatt tcaccacagg ggcttaatgt acctgtctta aattctaagg ttttaactcg
      901 ctttgtcaag catagacccc aaaaatttag ccaatgtctg taactcaatc tgtccatgtg
      961 tgggtgatga ggtacagtga cgctagcaca catcggaaaa acgctattac taggggaact
     1021 gaacagagta gcggacgcaa tgagtagtca tttaattggc ggttatgagc gtgttcaggc
     1081 ggtgctatca atcgtaatca taacagtggc agcttgatac agtgatgtca tccctgatgc
     1141 gaaagcgacc gaccgacggt acatcgaatg ggaatacttt agggtgattt ttaagaatcg
     1201 ctctagggtg agtatttccc attcagctct gctctctccc tctggtactt taatcaaaag
     1261 cactactaaa catatgtttt taaataaaaa atattgatat agagataata ttagtaagaa
     1321 taattaaaca attgaatata gataaatcat tgttaaataa agattaatta ttaaaatgaa
     1381 tgcatactta tatataaatc aatgatttaa aatatttgat aaagaaaact tttcaaaaaa
     1441 aatataattg agattgtgtc atttcggtca attcttaata tgttccacgc aagttttagc
     1501 tatggtgcta aacagaaatt tgctgaaaaa gaacttttca ctgaactggt taaaatgtaa
     1561 gcagcctgag agccgccaaa aattttaaaa acaaaccgcc ttaatcatct tcaaaaaata
     1621 cctctaaaac ctcaccattt gcgttttaag acccatattt catcctgccc ttatgttccc
     1681 atgctgatag ctataaagtg tctgtaatcg cttcctatga cgttctaggc tgttgataac
     1741 ttttggaaca acgcaaaatg ttaaaatcct gctgtgataa tgtcaacgga ctagtccaga
     1801 aaccgcagta catcggagcg cttttgaagc tgatgctcga gatctcggct tgaacgaatt
     1861 gttattatgc acccggtgcg gtatcaggcg gaccccacag aaactgtgct aattcttccg
     1921 gatcggtaaa accgctcaga tccagcggtg tttcttcaaa cacttcaaaa tcatgcagaa
     1981 aggtaaaggc cagcagttca cgggcaaaat cttcggtacg tttccactgt gcaccatcca
     2041 gcagggctgc cagaatttca cgatcaccac gaaatgcatt cagatgcagc tgaaccaggc
     2101 tataacggct atcaccggca taaacatcgg tgaaatcaac aataccggta acttcggttg
     2161 ctgccagatc aacaaaaatg ttggtgccat gcagatcacc atgaacaaaa cgcggttcgc
     2221 gacctgccag cagggtatca acatccggca gccaatcttc cagacggtcc agcagacgcg
     2281 gactcagata accccaacca cgatgatctt caacggttgc ggcacgacgt tcacgcagca
     2341 gttccggaaa aacttcgcta tgcggtgtca gaacggtatt accggtcagc ggaacacgat
     2401 gcagacgacc cagaacacgt cccagttcac gtgccagtgc cagcagtgca ttacgatcgg
     2461 tggtgccatc cattgcgcta cgccaggtgg tgccggtcat acggctcata accagatacg
     2521 gccacggcca tgcacctgta ccaggacgca gttcaccacg acctaacaga cgaggaaccg
     2581 gaaccggtgc atctgccaga actgcatagg cttcgctttc gcttgccagg ctttccggac
     2641 cacaccaatg ttcaccaaac agtttaataa caggatcagg ttcaccaacc agaaccggat
     2701 tggtgctttc acccggaaca cgcagaacag gcggaaccgg cagacccagt tcttccagtg
     2761 cacgacgtgc cagcggttcc caaaattctt gatcattgcg caggcttgca tagctatcat
     2821 cactatcaat acgatccagc agcagcaggc tttcttgggt catcgttgct gctccataac
     2881 atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt
     2941 acgaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt
     3001 tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga
     3061 accgaacagg cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcca
     3121 tgagcaaata ttatacgcaa ggcgacaagg ctcgagagtt caacagacag ccttcgttct
     3181 tatggccgtt caggaaaaag gccatccgtc aggatggcct tctgcttaat ttgatgcctg
     3241 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc aacgttcaaa
     3301 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     3361 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc agttccctac
     3421 tctcgcatgg ggagacccca cactaccatc ggcgctacgg cgtttcactt ctgagttcgg
     3481 catggggtca ggtgggacca ccgcgctact gccgccaggc aaattctgtt ttatcagacc
     3541 gcttctgcgt tctgatttaa tctgtatcag gctgaaaatc ttctctcatc cgccaaaaca
     3601 gccaagcttg catgcctgca ggtcgactct agaggatctt tgtagagctc atccatgcca
     3661 tgtgtaatcc cagcagcagt tacaaactca agaaggacca tgtggtcacg cttttcgttg
     3721 ggatctttcg aaaggacaga ttgtgtcgac aggtaatggt tgtctggtaa aaggacaggg
     3781 ccatcgccaa ttggagtatt ttgttgataa tggtctgcta gttgaacgga accatcttca
     3841 acgttgtggc gaattttgaa gttagctttg attccattct tttgtttgtc tgccgtgata
     3901 tatacattgt gtgagttaaa gttgtactcc agtttgtgtc caagaatgtt tccatcttct
     3961 ttaaaatcaa taccctttaa ctcgatacga ttaacaaggg tatcaccttc aaacttgact
     4021 tcagcacgcg tcttgtaggt cccgtcatct ttgaaagata tagtgcgttc ctgtacataa
     4081 ccttcgggca tggcactctt gaaaaagtca tgccgtttca tgtgatccgg ataacgggaa
     4141 aagcattgaa caccataggt cagagtagtg acaagtgttg gccacggaac aggtagtttt
     4201 ccagtagtgc aaataaattt aagggtgagt tttccgtttg tagcatcacc ttcaccctct
     4261 ccacggacag aaaatttgtg cccattaaca tcaccatcta attcaacaag aattgggaca
     4321 actccagtga aaagttcttc tcctttgctc atggtctgtt tcctgtgtgc tgctgatgtg
     4381 ctcagtatct tgttatccgc tcacattgtc aatattttat ctgattaatg agacgtcttt
     4441 acgacctgca cagccatacc acagcttccg atggctgcct gacgccagaa gcattggtgc
     4501 accgtgcagt cgatgataag ctgtcaaacc agatcaattc gcgctaactt acattaattg
     4561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     4681 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4741 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     4801 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     4861 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     4921 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     4981 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     5041 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     5101 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     5161 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     5221 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     5281 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     5341 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     5401 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     5461 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     5521 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     5581 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     5641 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     5701 cgaaaggttt tgcaccattc gatggtgtca acgtaaatgc atgccgcttc gccttccgtt
     5761 tgggcctaga attccggttt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
     5821 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
     5881 cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc
     5941 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
     6001 cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac
     6061 cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6121 aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc
     6181 ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct
     6241 ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc
     6301 tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa
     6361 gaagatcatc ttattaatca gataaaatat ttctagctag agtcgatctt cgccagcagg
     6421 gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag ccagagtttc
     6481 agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg gacgtgctca
     6541 tagtccacga cgcccgtgat tttgtagccc tggccgacgg ccagcaggta ggccgacagg
     6601 ctcatgccg
//
