LOCUS       sequence-382852-        3347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive T7 RNAP-mediated expression of LasI 3OC12-HSL synthase 
            for Las quorum sensing system. LasI sequence is codon optimized for 
            E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3347)
  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 3347)
  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/pT7 LasI
FEATURES             Location/Qualifiers
     source          1..3347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..46)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      42..89
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(134..152)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        220..324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(325..393,394..1185)
                     /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(543..562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1356..1944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1845..1864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2098..2115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2195..2214)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2573..2592
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2573..2591
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2623..2645
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2664..2681
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2694..2714
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
       61 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggataattct
      121 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
      181 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      241 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      301 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      361 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      421 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      481 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
      541 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
      601 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
      661 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
      721 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
      781 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
      841 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
      901 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
      961 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1021 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     1081 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1141 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1201 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1261 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1321 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1381 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1441 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1501 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1561 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1621 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1681 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1741 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1801 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1861 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1921 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1981 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2041 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2101 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     2161 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2221 taagccagta tacactccgc tatcgctacg cttcgttaat acagatgtag gtgttccaca
     2281 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     2341 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     2401 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     2461 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     2521 gcgcacccgt ggccaggacc caacgctgcc cgagatctcg atcccgcgaa attaatacga
     2581 ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta actttaagaa
     2641 ggagatatac catgtctggt tctcatcatc atcatcatca tggtagcagc ggcgaaaacc
     2701 tgtattttca gtccatgatc gtacaaattg gtcgccgcga agagttcgat aaaaaactgc
     2761 tgggcgagat gcacaagctg cgtgctcaag tgttcaagga gcgcaaaggc tgggacgtta
     2821 gtgtcatcga cgagatggaa atcgatggtt atgacgcact gagtccttat tacatgctga
     2881 tccaggaaga tactcctgaa gcccaggttt tcggttgctg gcgcattctg gataccactg
     2941 gcccgtacat gctgaagaac accttcccgg agctgctgca cggcaaggaa gcgccttgct
     3001 cgccgcacat ctgggaactg agccgtttcg ccatcaactc tggccagaaa ggctcgctgg
     3061 gcttttccga ctgtacgctg gaggcgatgc gcgcgctggc ccgctacagc ctgcagaacg
     3121 acatccagac gctggtgacg gtaaccaccg taggcgtgga gaagatgatg atccgtgccg
     3181 gcctggacgt atcgcgcttc ggtccgcacc tgaagatcgg catcgagcgc gcggtggccc
     3241 tgcgcatcga actgaatgcc aagacccaga tcgcgctgta cgggggcgtg ctggtggaac
     3301 agcgcctggc ggtttcatga agggtgatcc ggctgctaac aaagccc
//
