LOCUS       sequence-416352-        6080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-BIB H20A  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: pJS124
FEATURES             Location/Qualifiers
     source          1..6080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1007..1024)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1177..1263
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1355..1382
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1401..1492
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1493..1561,1562..2353)
                     /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(1711..1730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2395..2850
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2482..2501)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2692..2713
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2961..3549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3450..3469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3703..3720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3886..3908
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4046..4065
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4245..5123)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5150..5434
                     /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     5357..5376
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(5482..6080,1..109)
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
ORIGIN
        1 tctgagcacc cagagcaaac tgagcaaaga tccgaatgaa aaacgtgatc acatggtgct
       61 gctggaattt gttaccgcag caggtattac cggtggtagc ggtggttcag gtggtggtgg
      121 tagcaccctg gatctgcctc gtcgttttcc ttggccgacg ctgctgagcg tttgtattgc
      181 gggtgcagtt gttgcaggtc tgctgtatac cagcgttcat caggttattg aactgcctgc
      241 accggcacag cctgcagatt acaaaaccgg tggcaatggt aatggtggtc cgaccggtag
      301 cggtaatagc ggtggcggtg gctcaggcaa tggtggcggt aatggtaatt ctggtggtgg
      361 cagcggtaca ggtggtaatg gcagcgcagg taacggcaat ggcggtccta ccggctcagg
      421 taattcaggc ggaggcggtt caggtaatgg cggaggcaat ggcaacagcg gtggtggaag
      481 tggcactggt ggcaacggtt ctgcaggcaa cggtaacggt ggccctacag gttctggtaa
      541 cagtggtggc ggaggtagcg gcaacggtgg tggtaacgga aatagtggcg gtggttctgg
      601 caccggtgga aatggcagtg cacctggcac cggcaccgca gcaaccagca aaccggttac
      661 cagcgtggca agcggtccgc gtgcactgag ccgtaatcag ccgcagtatc cggcacgtgc
      721 acaggcactg cgtattgaag gtcaggttaa agtgaaattt gatgtgacac cggatggtcg
      781 tgttgataat gttcagattc tgagcgcaaa accggcaaat atgtttgaac gtgaagtgaa
      841 aaatgcaatg cgtcgttggc gttatgaacc gggtaaacct ggtagtggta ttgttgtgaa
      901 tatcctgttt aaaatcaacg gcaccaccga aatccagtaa atcctctaga gtcgacctgc
      961 aggcatgcaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
     1021 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
     1081 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
     1141 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     1201 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     1261 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
     1321 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
     1381 tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct
     1441 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1501 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1561 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1621 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1681 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     1741 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1801 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1861 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1921 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1981 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2041 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2101 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2161 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2221 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2281 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2341 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc
     2401 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2461 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2521 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2581 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     2641 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2701 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     2761 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2821 attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat
     2881 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2941 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3001 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3061 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3121 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3181 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3241 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3301 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3361 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3421 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3481 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3541 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3601 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3661 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3721 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3781 cgcatagggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3841 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3901 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gaaggagatg gcgcccaaca
     3961 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     4021 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     4081 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgc tcatgtttga
     4141 cagcttatca tcgatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc
     4201 ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg
     4261 gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg
     4321 cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg
     4381 gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc
     4441 tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac
     4501 ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga
     4561 tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg
     4621 ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc
     4681 gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc
     4741 ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta
     4801 agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga
     4861 gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc
     4921 ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga
     4981 ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg
     5041 gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg
     5101 ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat
     5161 tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc
     5221 aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca
     5281 aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac
     5341 ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga
     5401 cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc tagcaggagg
     5461 aattcaccat ggtacccggg gatggatagc accgaaagcc tgtttaccgg tgttgttccg
     5521 attctggttg aactggatgg tgatgttaat ggccacaaat tttcagttcg tggtgaaggc
     5581 gaaggtgatg caaccaatgg taaactgacc ctgaaattta tctgtaccac cggcaaactg
     5641 ccggttccgt ggccgacact ggttaccaca ctgacctatg gtgttcagtg ttttagccgt
     5701 tatccggatc atatgaaaca gcacgatttc tttaaaagcg caatgccgga aggttatgtt
     5761 caagaacgta ccatcacgtt taaagatgat ggcacctata aaacccgtgc cgaagttaaa
     5821 tttgaaggtg ataccctggt gaatcgcatt gaactgaaag gcattgattt caaagaagat
     5881 ggcaatatcc tgggccacaa actggaatat aattataaca gccacaacgt gtatatcacc
     5941 gcagacaaac agaaaaatgg catcaaagcc aactttaaga ttcggcataa tgtggaagat
     6001 ggtagcgttc agctggcaga tcattatcag cagaataccc cgattggtga tggtccggtt
     6061 ctgctgccgg ataatcatta
//
