LOCUS       sequence-414350-        6579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-AABB expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6579)
  AUTHORS   Williams-Jones DP, Webby MN, Press CE, Gradon JM, Armstrong SR, 
            Szczepaniak J, Kleanthous C
  TITLE     Tunable force transduction through the Escherichia coli cell 
            envelope.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Nov 21;120(47):e2306707120. doi: 
            10.1073/pnas.2306707120. Epub 2023 Nov 16.
  PUBMED    37972066
REFERENCE   2  (bases 1 to 6579)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-11-21"; volume: "120(47):e2306707120. 
            doi"; pages: " 10.1073/pnas.2306707120. Epub 2023 Nov 16"
COMMENT     Sequence Label: pDWJ14
FEATURES             Location/Qualifiers
     source          1..6579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1362..2075
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1362..2075
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1392..1420)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2015..2035
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2076..2105
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     primer_bind     complement(3437..3454)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3607..3693
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3785..3812
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3831..3922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3923..3991,3992..4783)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4141..4160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4825..5280
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4912..4931)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5122..5143
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5391..5979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5880..5899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6133..6150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6316..6338
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6476..6495
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg ttttttgggc tagcaggagg aattcaccat
     1321 ggtacccggg gatcctctag atttaagaag gagatataca tatgagtaaa ggagaagaac
     1381 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
     1441 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
     1501 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ttcgcgtatg
     1561 gtcttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt ttcaagagtg
     1621 ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca
     1681 agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
     1741 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
     1801 cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt aacttcaaaa
     1861 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
     1921 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     1981 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
     2041 ctgggattac acatggcatg gatgaactat acaaagctag catgactggt ggacagcaaa
     2101 tgggtcgcgg atccgtgtca aaggcaaccg aacaaaacga caagctcaag cgggcgataa
     2161 ttatttcagc agtgctgcat gtcatcttat ttgcggcgct gatctggagt tcgttcgatg
     2221 agaatataga agcttcagcc ggaggcggcg gtggttcgtc catcgacgct gtcatggttg
     2281 attcaggtgc ggtagttgag cagtacaaac gcatgcaaag ccaggaatca agcgcgaagc
     2341 gttctgatga acagcgcaag atgaaggaac agcaggctgc tgaagaactg cgtgagaaac
     2401 aagcggctga acaggaacgc ctgaagcaac ttgagaaaga gcggttagcg gctcaggagc
     2461 agaaaaagca ggctgaagaa gccgcaaaac aggccgagtt aaagcagaag caagctgaag
     2521 aggcggcagc gaaagcggcg gcagatgcta aagcgaaggc cgaagcagat gctaaagctg
     2581 cggaagaagc agcgaagaaa gcggctgcag acgcaaagaa aaaagcagaa gcagaagccg
     2641 ccaaagccgc agccgaagcg cagaaaaaag ccgaggcagc cgctgcggca ctgaagaaga
     2701 aagcggaagc ggcagaagca gctgcagctg aagcaagaaa gaaagcggca actgaagctg
     2761 ctgaaaaagc caaacctgcg cctgcgcagc cgatttctgt cacgatggtt acgcctgctg
     2821 atctcgaacc gccacaagcc gttcagccgc caccggagcc ggtggtagag ccagaaccgg
     2881 aacctgagcc gatccccgaa ccgccaaaag aagcaccggt ggtcattgaa aagccgaagc
     2941 cgaaacctaa gccaaaaccg aagccggtga aaaaggtaca ggagcagcca aaacgcgatg
     3001 tcaaacccgt agagtcgcgt ccggcatcac cgtttgaaaa tacggcaccg gcacgcctga
     3061 catcaagtac agcaacggct gcaaccagca agccggttac cagtgtggct tcaggaccac
     3121 gcgcattaag ccgtaatcag ccgcagtatc cggcacgagc acaggcattg cgcattgaag
     3181 ggcaggttaa agttaaattt gacgttacgc cggatggtcg cgtggataac gtacaaatcc
     3241 tctcagccaa gcctgcgaac atgtttgagc gtgaggtgaa aaatgcgatg cgcagatggc
     3301 gttatgagcc gggtaagcca ggcagtggga ttgtggtgaa tatcctgttt aaaattaacg
     3361 gcaccaccga aattcagtaa gtcgacctgc aggcatgcaa gcttggctgt tttggcggat
     3421 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     3481 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     3541 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     3601 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3661 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     3721 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     3781 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     3841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3901 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3961 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4021 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4081 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4141 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     4201 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4261 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4321 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4381 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4441 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4501 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4561 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4621 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4681 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4741 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4801 ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4861 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4921 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4981 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5041 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5101 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5161 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5221 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5281 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5341 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5401 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5461 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5521 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5581 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5641 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5701 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5761 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5821 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5881 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5941 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6001 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6061 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6121 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     6181 ctccttacgc atctgtgcgg tatttcacac cgcatagggt catggctgcg ccccgacacc
     6241 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6301 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6361 gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     6421 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     6481 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     6541 gtccggcgta gaggatctgc tcatgtttga cagcttatc
//
