LOCUS       sequence-381997-        3636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of eGFP by the transcription factor complex 
            LuxR-3OC6 homoserine lactone (HSL)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3636)
  AUTHORS   Gonzales DT, Suraritdechachai S, Zechner C, Tang T-YD
  TITLE     Bidirectional communication between droplet interface bilayers 
            driven by cell-free quorum sensing gene circuits
  JOURNAL   ChemSystemsChem 2023
REFERENCE   2  (bases 1 to 3636)
  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: 
            "ChemSystemsChem 2023"
COMMENT     Sequence Label: pEXP5-NT/pLux eGFP
FEATURES             Location/Qualifiers
     source          1..3636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             172..194
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             213..230
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             243..263
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             285..299
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             join(318..320,321..323,324..1034)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(318..320,321..323,324..1034)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(363..384)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(624..643)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     971..992
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1116..1134)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1130..1177
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1222..1240)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1308..1412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1413..1481,1482..2273)
                     /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(1631..1650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2444..3032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2933..2952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3186..3203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3283..3302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cccgagatct gagctggcgt acgcgttgaa cacttcacag atgataggga ttcgggtaaa
       61 gagcgtgtca ttgggggctt atacaggcgt tacaattact taacataagc acctgtagga
      121 tcgtacaggt ttacgcaaga aaatggtttg ttatagtcgt ctagaaataa ttttgtttaa
      181 ctttaagaag gagatatacc atgtctggtt ctcatcatca tcatcatcat ggtagcagcg
      241 gcgaaaacct gtattttcag tcccttgtgg gtaccggttc gaatgatgac gacgacaaga
      301 gtccggatcc caattggatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      361 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      421 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      481 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
      541 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      601 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      661 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
      721 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
      781 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
      841 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
      901 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
      961 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1021 acgagctgta caagtccgga ctcagatctc gagtctggta aagggtgatc cggctgctaa
     1081 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1141 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1201 ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1261 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1321 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1381 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1441 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1501 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1561 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1621 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1681 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1741 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1801 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1861 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1921 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1981 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2041 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2101 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2161 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2221 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2281 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2341 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2581 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3001 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3061 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3121 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3181 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3241 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     3301 cgcatagtta agccagtata cactccgcta tcgctacgct tcgttaatac agatgtaggt
     3361 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3421 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3481 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3541 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     3601 acgatcatgc gcacccgtgg ccaggaccca acgctg
//
