LOCUS       sequence-414366-        6036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-ABB expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6036)
  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 6036)
  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: pDWJ17
FEATURES             Location/Qualifiers
     source          1..6036
                     /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..1284
                     /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)"
     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(2894..2911)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3064..3150
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3242..3269
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3288..3379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3380..3448,3449..4240)
                     /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(3598..3617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4282..4737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4369..4388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4579..4600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4848..5436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5337..5356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5590..5607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5773..5795
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5933..5952
                     /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 agcggaacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt ttatccataa gattagcgga tcctacctga cgctttttat
     1261 cgcaactctc tactgtttct ccatacccgt 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 ggaggcggca tttctgtcac gatggttacg cctgctgatc
     2281 tcgaaccgcc acaagccgtt cagccgccac cggagccggt ggtagagcca gaaccggaac
     2341 ctgagccgat ccccgaaccg ccaaaagaag caccggtggt cattgaaaag ccgaagccga
     2401 aacctaagcc aaaaccgaag ccggtgaaaa aggtacagga gcagccaaaa cgcgatgtca
     2461 aacccgtaga gtcgcgtccg gcatcaccgt ttgaaaatac ggcaccggca cgcctgacat
     2521 caagtacagc aacggctgca accagcaagc cggttaccag tgtggcttca ggaccacgcg
     2581 cattaagccg taatcagccg cagtatccgg cacgagcaca ggcattgcgc attgaagggc
     2641 aggttaaagt taaatttgac gttacgccgg atggtcgcgt ggataacgta caaatcctct
     2701 cagccaagcc tgcgaacatg tttgagcgtg aggtgaaaaa tgcgatgcgc agatggcgtt
     2761 atgagccggg taagccaggc agtgggattg tggtgaatat cctgtttaaa attaacggca
     2821 ccaccgaaat tcagtaagtc gacctgcagg catgcaagct tggctgtttt ggcggatgag
     2881 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     2941 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     3001 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     3061 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3121 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     3181 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     3241 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     3301 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3361 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3421 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3481 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3541 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3601 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     3661 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3721 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3781 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3841 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3901 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3961 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4021 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4081 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4141 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4201 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4261 atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4321 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4381 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4441 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4501 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4561 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4621 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4681 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
     4741 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4801 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4861 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4921 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4981 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5041 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5101 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5161 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5221 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5281 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5341 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5401 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5461 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5521 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5581 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     5641 cttacgcatc tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc cgacacccgc
     5701 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5761 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5821 cgaggcagaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     5881 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     5941 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6001 cggcgtagag gatctgctca tgtttgacag cttatc
//
