LOCUS       sequence-439071-        6411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with GFP expressed under PabstBR promoter, aprR 
            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 6411)
  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 6411)
  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: pJMP3666
FEATURES             Location/Qualifiers
     source          1..6411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      31..117
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      209..236
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(352..357,358..374,375..380)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             569..1369
                     /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            2991..3100
                     /note="incP origin of transfer"
     rep_origin      3483..4028
                     /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     3929..3948
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4114..4191
                     /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             4192..5274
                     /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(4211..4230)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             join(5491..5493,5494..6204)
                     /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             5491..6204
                     /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(5521..5549)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6144..6164
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6272..6289)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
       61 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
      121 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
      181 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttcct
      241 gaacggccat aagaacgaag gctgtctgtt gaactctcga gccttgtcgc cttgcgtata
      301 atatttgctc atggacgcac accgtggaaa cggatgaagg cacgaaccca gttgacataa
      361 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
      421 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
      481 tgttttttcg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
      541 cgatgtttga tgttatggag cagcaacgat gtcatcagcg gtggagtgca atgtcgtgca
      601 atacgaatgg cgaaaagccg agctgatcgg tcagcttctc aaccttgggg ttacccccgg
      661 cggtgtgctg ctggtccaca gctccttccg tagcgtccgg cccctcgaag atgggccact
      721 tggactgatc gaggccctgc gtgctgcact gggtccggga gggacgctcg tcatgccatc
      781 gtggtcaggt ctggacgacg agccgttcga tcctgccacg tcgcccgtta caccggacct
      841 tggagttgtc tctgacacat tctggcgtct gccaaatgta aagcgcagcg cccatccatt
      901 tgcctttgcg gcagcggggc cacaggcaga gcagatcatc tctgatccat tgcccctgcc
      961 acctcactcg cctgcaagcc cggtcgcccg tgtccatgaa ctcgatgggc aggtacttct
     1021 cctcggcgtg ggacacgatg ccaacacgac gctgcatctt gccgagttga tggcaaaggt
     1081 tccctatggg gtgccgagac actgcaccat tcttcaggat ggcaagttgg tacgtgtcga
     1141 ttatctggag aatgaccact gctgtgagcg ctttgccttg gcggacaggt ggctcaagga
     1201 gaacagcctt cagaaggaag gtccagtcgg tcatgccttt gctcggttga tccgctcccg
     1261 cgacattgtg gcgacagccc tgggtcaact gggccgagat ccgttgatct tcctgcatcc
     1321 gccagaggcg ggatgcgaag aatgcgatgc cgctcgccag tcgattggct aacaattcgt
     1381 tcaagccgag atctcgagca tcagcttcaa aagcgctccg atgtactgcg gtttctggac
     1441 tagtccgttg acattatcac agcaggattt taacattttg cgttgttcca aaagttatca
     1501 acagcctaga acgtcatagg aagcgattac agacacttta tagctatcag catgggaaca
     1561 taagggcagg atgaaatatg ggtcttaaaa cgcaaatggt gaggttttag aggtattttt
     1621 tgaagatgat taaggcggtt tgtttttaaa atttttggcg gctctcaggc tgcttacatt
     1681 ttaaccagtt cagtgaaaag ttctttttca gcaaatttct gtttagcacc atagctaaaa
     1741 cttgcgtgga acatattaag aattgaccga aatgacacaa tctcaattat attttttttg
     1801 aaaagttttc tttatcaaat attttaaatc attgatttat atataagtat gcattcattt
     1861 taataattaa tctttattta acaatgattt atctatattc aattgtttaa ttattcttac
     1921 taatattatc tctatatcaa tattttttat ttaaaaacat atgtttagta gtgcttttga
     1981 ttaaagtacc agagggagag agcagagctg aatgggaaat actcacccta gagcgattct
     2041 taaaaatcac cctaaagtat tcccattcga tgtaccgtcg gtcggtcgct ttcgcatcag
     2101 ggatgacatc actgtatcaa gctgccactg ttatgattac gattgatagc accgcctgaa
     2161 cacgctcata accgccaatt aaatgactac tcattgcgtc cgctactctg ttcagttccc
     2221 ctagtaatag cgtttttccg atgtgtgcta gcgtcactgt acctcatcac ccacacatgg
     2281 acagattgag ttacagacat tggctaaatt tttggggtct atgcttgaca aagcgagtta
     2341 aaaccttaga atttaagaca ggtacattaa gcccctgtgg tgaaatcatt agcggtcgtc
     2401 aaaccttaat gctttcgatt gccgatatgt cagtatcgaa gatctgtacc ccataaatta
     2461 cggtaaagcc ccaagcaatt gcaaggggct tttatctttt ttaacaaaaa aaatttataa
     2521 atcaggattt tataacacta aataatccaa agtacatgag taagtcatga ccactcctgc
     2581 gatgtgtgta gcactctgag tatccgtatt atatcagtga tgtcatatac aaccacataa
     2641 tgcggatgta tcactaactc cctagtatct ttctgtctgc ctgtgcgccc catcagtcga
     2701 ttatcaacaa gtaaatctgt cttttcttca attaaatcat caatttcaat agctgctata
     2761 gggttgcgtt cttcaagata atcatagata tttctacgat cttggcttgc agcccttatg
     2821 tcctcaggcg cgcacaaaac agcagggaag cagcgctttt ccgctgcata accctgcttc
     2881 ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt
     2941 tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata
     3001 ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg
     3061 gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg cgtaacagat
     3121 gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc acctatcaag
     3181 gtgtactgcc ttccagacga acgaagagcg attgaggaaa aggcggcggc ggccggcatg
     3241 agcctgtcgg cctacctgct ggccgtcggc cagggctaca aaatcacggg cgtcgtggac
     3301 tatgagcacg tccgcgagct ggcccgcatc aatggcgacc tgggccgcct gggcggcctg
     3361 ctgaaactct ggctcaccga cgacccgcgc acggcgcggt tcggtgatgc cacgatcctc
     3421 gccctgctgg cgaagatcga ctctagctag aaatatttta tctgattaat aagatgatct
     3481 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     3541 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     3601 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     3661 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     3721 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     3781 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     3841 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     3901 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     3961 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     4021 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaaaccgga attctaggcc
     4081 caaacggaag gcgaagcggc atgcatttac gttgacacca tcgaatggtg caaaaccttt
     4141 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     4201 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     4261 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     4321 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     4381 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     4441 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     4501 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     4561 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     4621 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     4681 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     4741 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     4801 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     4861 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     4921 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     4981 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     5041 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     5101 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     5161 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5221 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     5281 cgcaattaat gtaagttagc gcgaattgat ctggtttgac agcttatcat cgactgcacg
     5341 gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc
     5401 gtaaagacgt ctcattaatc agataaaata ttgacaatgt gagcggataa caagatactg
     5461 agcacatcag cagcacacag gaaacagacc atgagcaaag gagaagaact tttcactgga
     5521 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt
     5581 ggagagggtg aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact
     5641 ggaaaactac ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc
     5701 ttttcccgtt atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa
     5761 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct
     5821 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt
     5881 aaagaagatg gaaacattct tggacacaaa ctggagtaca actttaactc acacaatgta
     5941 tatatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac
     6001 gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
     6061 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat
     6121 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
     6181 catggcatgg atgagctcta caaagatcct ctagagtcga cctgcaggca tgcaagcttg
     6241 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
     6301 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     6361 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct c
//
