LOCUS       sequence-382851-        3460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive T7 RNAP-mediated expression of LasR transcription 
            factor for Las quorum sensing system. LasR sequence is codon 
            optimized for E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3460)
  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 3460)
  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 LasR
FEATURES             Location/Qualifiers
     source          1..3460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..42)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      38..85
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(130..148)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        216..320
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(321..389,390..1181)
                     /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(539..558)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1352..1940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1841..1860
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2094..2111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2191..2210)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2569..2588
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2569..2587
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2619..2641
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2660..2677
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2690..2710
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
       61 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat aattcttgaa
      121 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
      181 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      241 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      301 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      361 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      421 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      481 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      541 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
      601 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      661 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      721 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      781 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      841 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      901 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      961 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1021 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1081 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1141 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1201 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1261 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1321 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1381 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1441 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1501 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1561 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1621 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1681 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1741 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1801 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1861 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1921 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1981 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2041 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2101 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     2161 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2221 ccagtataca ctccgctatc gctacgcttc gttaatacag atgtaggtgt tccacagggt
     2281 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     2341 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     2401 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     2461 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     2521 acccgtggcc aggacccaac gctgcccgag atctcgatcc cgcgaaatta atacgactca
     2581 ctatagggag accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag
     2641 atataccatg tctggttctc atcatcatca tcatcatggt agcagcggcg aaaacctgta
     2701 ttttcagtcc atggccttgg ttgacggttt tcttgagctg gaacgctcaa gtggaaaatt
     2761 ggagtggagc gccatcctcc agaagatggc gagcgacctt ggattctcga agatcctgtt
     2821 cggcctgttg cctaaggaca gccaggacta cgagaacgcc ttcatcgtcg gcaactaccc
     2881 ggccgcctgg cgcgagcatt acgaccgggc tggctacgcg cgggtcgacc cgacggtcag
     2941 tcactgtacc cagagcgtac tgccgatttt ctgggaaccg tccatctacc agacgcgaaa
     3001 gcagcacgag ttcttcgagg aagcctcggc cgccggcctg gtgtatgggc tgaccatgcc
     3061 gctgcatggt gctcgcggcg aactcggcgc gctgagcctc agcgtggaag cggaaaaccg
     3121 ggccgaggcc aaccgtttca tagagtcggt cctgccgacc ctgtggatgc tcaaggacta
     3181 cgcactgcaa agcggtgccg gactggcctt cgaacatccg gtcagcaaac cggtggttct
     3241 gaccagccgg gagaaggaag tgttgcagtg gtgcgccatc ggcaagacca gttgggagat
     3301 atcggttatc tgcaactgct cggaagccaa tgtgaacttc catatgggaa atattcggcg
     3361 gaagttcggt gtgacctccc gccgcgtagc ggccattatg gccgttaatt tgggtcttat
     3421 tactctctaa aggtgatccg gctgctaaca aagcccgaaa
//
