LOCUS       sequence-414341-        6332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-ABBA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6332)
  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 6332)
  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: pJS109
FEATURES             Location/Qualifiers
     source          1..6332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..83
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     249..271
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     409..428
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(608..1486)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1513..1797
                     /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     1720..1739
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1855..2568
                     /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             1855..2568
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1885..1913)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2508..2528
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2569..2598
                     /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(3702..3719)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3872..3958
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4050..4077
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4096..4187
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4188..4256,4257..5048)
                     /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(4406..4425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5090..5545
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5177..5196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5387..5408
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5656..6244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6145..6164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
       61 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
      121 cgcatctgtg cggtatttca caccgcatag ggtcatggct gcgccccgac acccgccaac
      181 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
      241 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
      301 gcagaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
      361 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
      421 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
      481 gtagaggatc tgctcatgtt tgacagctta tcatcgatgc ataatgtgcc tgtcaaatgg
      541 acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg
      601 ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
      661 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
      721 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
      781 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
      841 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
      901 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
      961 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     1021 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     1081 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     1141 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     1201 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     1261 gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca
     1321 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     1381 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     1441 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
     1501 cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct
     1561 tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac
     1621 aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa
     1681 aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
     1741 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc
     1801 cgtttttttg ggctagcagg aggaattcac catggtaccc ggggatcctc tagaatgagt
     1861 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     1921 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     1981 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     2041 actttcgcgt atggtcttca atgctttgcg agatacccag atcatatgaa acagcatgac
     2101 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     2161 gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     2221 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     2281 tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa
     2341 gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     2401 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     2461 acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     2521 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaagc tagcatgact
     2581 ggtggacagc aaatgggtcg cggatccgtg tcaaaggcaa ccgaacaaaa cgacaagctc
     2641 aagcgggcga taattatttc agcagtgctg catgtcatct tatttgcggc gctgatctgg
     2701 agttcgttcg atgagaatat agaagcttca gccggaggcg gctctgtcac gatggttacg
     2761 cctgctgatc tcgaaccgcc acaagccgtt cagccgccac cggagccggt ggtagagcca
     2821 gaaccggaac ctgagccgat ccccgaaccg ccaaaagaag caccggtggt cattgaaaag
     2881 ccgaagccga aacctaagcc aaaaccgaag ccggtgaaaa aggtacagga gcagccaaaa
     2941 cgcgatgtca aacccgtaga gtcgcgtccg gcatcaccgt ttgaaaatac ggcaccggca
     3001 cgcctgacat caagtacagc aacgatttct gtcacgatgg ttacgcctgc tgatctcgaa
     3061 ccgccacaag ccgttcagcc gccaccggag ccggtggtag agccagaacc ggaacctgag
     3121 ccgatccccg aaccgccaaa agaagcaccg gtggtcattg aaaagccgaa gccgaaacct
     3181 aagccaaaac cgaagccggt gaaaaaggta caggagcagc caaaacgcga tgtcaaaccc
     3241 gtagagtcgc gtccggcatc accgtttgaa aatacggcac cggcacgcct gacatcaagt
     3301 acagcaacgg ggggaggggc gaaagggaac aatgcttcgc ctgccgggag tggtaatact
     3361 aaaaacaatg gcgcatcagg ggccgatatc aataactatg ccgggcagat taaatctgct
     3421 atcgaaagta agttctatga cgcatcgtcc tatgcaggca aaacctgtac gctgcgcata
     3481 aaactggcac ccgatggtat gttactggat atcaaacctg aaggtggcga tcccgcactt
     3541 tgtcaggctg cgttggcagc agctaaactt gcgaagatcc cgaaaccacc aagccaggca
     3601 gtatatgaag tgttcaaaaa cgcgccattg gacttcaaac cgtaagtcga cctgcaggca
     3661 tgcaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     3721 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     3781 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     3841 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     3901 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     3961 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     4021 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     4081 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4141 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4201 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4261 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4321 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4381 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     4441 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4501 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4561 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4621 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4681 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4741 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4801 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4861 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4921 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4981 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5041 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta cgcgccctgt
     5101 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5161 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5221 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5281 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     5341 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5401 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     5461 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5521 aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt ttgataatct
     5581 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5761 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     5821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5881 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5941 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6001 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6061 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6121 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6181 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6241 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6301 catgttcttt cctgcgttat cccctgattc tg
//
