LOCUS       sequence-414398-        6080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-BIB expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6080)
  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 6080)
  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: pDWJ27
FEATURES             Location/Qualifiers
     source          1..6080
                     /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             1333..2040
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(2938..2955)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3108..3194
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3286..3313
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3332..3423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3424..3492,3493..4284)
                     /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(3642..3661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4326..4781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4413..4432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4623..4644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4892..5480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5381..5400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5634..5651
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5817..5839
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5977..5996
                     /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 tttttttggg ctagcaggag gaattcacca
     1321 tggtacccgg ggatggatag caccgaaagc ctgtttaccg gtgttgttcc gattctggtt
     1381 gaactggatg gtgatgttaa tggccacaaa ttttcagttc gtggtgaagg cgaaggtgat
     1441 gcaaccaatg gtaaactgac cctgaaattt atctgtacca ccggcaaact gccggttccg
     1501 tggccgacac tggttaccac actgacctat ggtgttcagt gttttagccg ttatccggat
     1561 catatgaaac agcacgattt ctttaaaagc gcaatgccgg aaggttatgt tcaagaacgt
     1621 accatcacgt ttaaagatga tggcacctat aaaacccgtg ccgaagttaa atttgaaggt
     1681 gataccctgg tgaatcgcat tgaactgaaa ggcattgatt tcaaagaaga tggcaatatc
     1741 ctgggccaca aactggaata taattataac agccacaacg tgtatatcac cgcagacaaa
     1801 cagaaaaatg gcatcaaagc caactttaag attcggcata atgtggaaga tggtagcgtt
     1861 cagctggcag atcattatca gcagaatacc ccgattggtg atggtccggt tctgctgccg
     1921 gataatcatt atctgagcac ccagagcaaa ctgagcaaag atccgaatga aaaacgtgat
     1981 cacatggtgc tgctggaatt tgttaccgca gcaggtatta ccggtggtag cggtggttca
     2041 ggtggtggtg gtagcaccct ggatctgcct cgtcgttttc cttggccgac gctgctgagc
     2101 gtttgtattc atggtgcagt tgttgcaggt ctgctgtata ccagcgttca tcaggttatt
     2161 gaactgcctg caccggcaca gcctgcagat tacaaaaccg gtggcaatgg taatggtggt
     2221 ccgaccggta gcggtaatag cggtggcggt ggctcaggca atggtggcgg taatggtaat
     2281 tctggtggtg gcagcggtac aggtggtaat ggcagcgcag gtaacggcaa tggcggtcct
     2341 accggctcag gtaattcagg cggaggcggt tcaggtaatg gcggaggcaa tggcaacagc
     2401 ggtggtggaa gtggcactgg tggcaacggt tctgcaggca acggtaacgg tggccctaca
     2461 ggttctggta acagtggtgg cggaggtagc ggcaacggtg gtggtaacgg aaatagtggc
     2521 ggtggttctg gcaccggtgg aaatggcagt gcacctggca ccggcaccgc agcaaccagc
     2581 aaaccggtta ccagcgtggc aagcggtccg cgtgcactga gccgtaatca gccgcagtat
     2641 ccggcacgtg cacaggcact gcgtattgaa ggtcaggtta aagtgaaatt tgatgtgaca
     2701 ccggatggtc gtgttgataa tgttcagatt ctgagcgcaa aaccggcaaa tatgtttgaa
     2761 cgtgaagtga aaaatgcaat gcgtcgttgg cgttatgaac cgggtaaacc tggtagtggt
     2821 attgttgtga atatcctgtt taaaatcaac ggcaccaccg aaatccagta aatcctctag
     2881 agtcgacctg caggcatgca agcttggctg ttttggcgga tgagagaaga ttttcagcct
     2941 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     3001 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     3061 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     3121 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     3181 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     3241 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     3301 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
     3361 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3421 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3481 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3541 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3601 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3661 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3721 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3781 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3841 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3901 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3961 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4021 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4081 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4141 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4201 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4261 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4321 gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4381 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4441 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4501 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     4561 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4621 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     4681 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4741 atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga
     4801 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4861 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4921 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4981 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5041 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5101 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5161 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5221 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5281 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5341 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5401 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5461 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5521 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5581 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5641 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5701 gtatttcaca ccgcataggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     5761 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5821 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agaaggagat
     5881 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     5941 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     6001 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg
     6061 ctcatgtttg acagcttatc
//
