LOCUS       sequence-251602-        4028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the transcriptional activator LuxR and GFP. LuxR has an 
            pLlacO-1 promoter and the GFP has a pLux promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4028)
  AUTHORS   Schwarz-Schilling M, Aufinger L, Muckl A, Simmel FC
  TITLE     Chemical communication between bacteria and cell-free gene 
            expression systems within linear chains of emulsion droplets.
  JOURNAL   Integr Biol (Camb). 2016 Apr 18;8(4):564-70. doi: 
            10.1039/c5ib00301f. Epub 2016 Jan 18.
  PUBMED    26778746
REFERENCE   2  (bases 1 to 4028)
  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"; doi: 
            "10.1039/c5ib00301f"; journalName: "Integr Biol (Camb)"; date: 
            "2016-04-18- 18"; volume: "8"; issue: "4"; pages: "564-70"
COMMENT     Sequence Label: ""AND-gate"" receiver plasmid
FEATURES             Location/Qualifiers
     source          1..4028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    27..43
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             64..75
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      879..950
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      966..993
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1090..1803
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1120..1148)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1743..1763
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1826..1897
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1913..1940
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1947..1967
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1973..1990)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2144..2732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2224..2243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2903..3694,3695..3763))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3526..3545
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3764..3868)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3936..3954
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3958..4001)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4004..4025
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ataaatgtga gcggataaca ttgacattgt gagcggataa caagatactg agcactacta
       61 gagaaagagg agaaatacta gatgaaaaac ataaatgccg acgacacata cagaataatt
      121 aataaaatta aagcttgtag aagcaataat gatattaatc aatgcttatc tgatatgact
      181 aaaatggtac attgtgaata ttatttactc gcgatcattt atcctcattc tatggttaaa
      241 tctgatattt caatcctaga taattaccct aaaaaatgga ggcaatatta tgatgacgct
      301 aatttaataa aatatgatcc tatagtagat tattctaact ccaatcattc accaattaat
      361 tggaatatat ttgaaaacaa tgctgtaaat aaaaaatctc caaatgtaat taaagaagcg
      421 aaaacatcag gtcttatcac tgggtttagt ttccctattc atacggctaa caatggcttc
      481 ggaatgctta gttttgcaca ttcagaaaaa gacaactata tagatagttt atttttacat
      541 gcgtgtatga acataccatt aattgttcct tctctagttg ataattatcg aaaaataaat
      601 atagcaaata ataaatcaaa caacgattta accaaaagag aaaaagaatg tttagcgtgg
      661 gcatgcgaag gaaaaagctc ttgggatatt tcaaaaatat taggttgcag tgagcgtact
      721 gtcactttcc atttaaccaa tgcgcaaatg aaactcaata caacaaaccg ctgccaaagt
      781 atttctaaag caattttaac aggagcaatt gattgcccat actttaaaaa ttaataacac
      841 tgatagtgct agtgtagatc actactagag ccaggcatca aataaaacga aaggctcagt
      901 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
      961 acactggctc accttcgggt gggcctttct gcgtttatat actagagacc tgtaggatcg
     1021 tacaggttta cgcaagaaaa tggtttgtta tagtcgaata aatactagag tcacacagga
     1081 aagtactaga tgcgtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     1141 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
     1201 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
     1261 ccaacacttg tcactacttt cggttatggt gttcaatgct ttgcgagata cccagatcat
     1321 atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
     1381 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
     1441 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     1501 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     1561 aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     1621 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     1681 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac
     1741 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     1801 aaataataat actagagcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     1861 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     1921 ttcgggtggg cctttctgcg tttatatact agtagcggcc gctgcaggct tcctcgctca
     1981 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2041 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2101 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccac aggctccgcc
     2161 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2221 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2281 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2341 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2401 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2461 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2521 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2581 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2641 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2701 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2761 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2821 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2881 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2941 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3001 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3061 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3121 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3181 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3241 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3301 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3361 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3421 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3481 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3541 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3601 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3661 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3721 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3781 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3841 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     3901 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt
     3961 tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg cggccgcttc
     4021 tagagctg
//
