LOCUS       sequence-382855-        3671 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Inducible expression of eGFP by the transcription factor complex 
            LasR-3OC12 homoserine lactone (HSL)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3671)
  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 3671)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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/pLas eGFP
FEATURES             Location/Qualifiers
     source          1..3671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             207..229
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             248..265
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             278..298
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             320..334
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             join(353..355,356..358,359..1069)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(353..355,356..358,359..1069)
                     /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(398..419)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(659..678)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1006..1027
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1151..1169)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1165..1212
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1257..1275)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1343..1447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1448..1516,1517..2308)
                     /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(1666..1685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2479..3067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2968..2987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3221..3238
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3318..3337)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cccgagatct gagctggcgt acgcgttgaa cacttcacag atgataggga ttcgggtaaa
       61 gagcgtgtca ttgggggctt atacaggcgt tgctgttccg ctgggcatgc ttcgagccta
      121 gcaagggtcc gggttcaccg aaatctatct catttgctag ttataaaatt atgaaatttg
      181 cgtaaattct tcagtctaga aataattttg tttaacttta agaaggagat ataccatgtc
      241 tggttctcat catcatcatc atcatggtag cagcggcgaa aacctgtatt ttcagtccct
      301 tgtgggtacc ggttcgaatg atgacgacga caagagtccg gatcccaatt ggatggtgag
      361 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      421 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      481 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      541 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
      601 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      661 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      721 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
      781 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
      841 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
      901 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
      961 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1021 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag
     1081 atctcgagtc tggtaaaggg tgatccggct gctaacaaag cccgaaagga agctgagttg
     1141 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1201 aggggttttt tgctgaaagg aggaactata tccggataat tcttgaagac gaaagggcct
     1261 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1321 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1381 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1441 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1501 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1561 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1621 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1681 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1741 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1801 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1861 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1921 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1981 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2041 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2101 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2161 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2221 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2281 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2341 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2401 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2461 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2521 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2581 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2641 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2701 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2761 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2821 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2881 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2941 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3001 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3061 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3121 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3181 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3241 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3301 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     3361 cgctatcgct acgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     3421 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     3481 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     3541 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     3601 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     3661 acccaacgct g
//
