LOCUS       sequence-244585-        3964 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 3964)
  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 3964)
  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..3964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    30..46
                     /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             67..78
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      882..953
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      969..996
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1093..1806
                     /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(1123..1151)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1746..1766
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    1810..1830
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1831..1888
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1912..1929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2083..2671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2163..2182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2842..3633,3634..3702))
                     /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     3465..3484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3703..3807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3875..3893
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3897..3940)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3943..3964
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ctgataaatg tgagcggata acattgacat tgtgagcgga taacaagata ctgagcacta
       61 ctagagaaag aggagaaata ctagatgaaa aacataaatg ccgacgacac atacagaata
      121 attaataaaa ttaaagcttg tagaagcaat aatgatatta atcaatgctt atctgatatg
      181 actaaaatgg tacattgtga atattattta ctcgcgatca tttatcctca ttctatggtt
      241 aaatctgata tttcaatcct agataattac cctaaaaaat ggaggcaata ttatgatgac
      301 gctaatttaa taaaatatga tcctatagta gattattcta actccaatca ttcaccaatt
      361 aattggaata tatttgaaaa caatgctgta aataaaaaat ctccaaatgt aattaaagaa
      421 gcgaaaacat caggtcttat cactgggttt agtttcccta ttcatacggc taacaatggc
      481 ttcggaatgc ttagttttgc acattcagaa aaagacaact atatagatag tttattttta
      541 catgcgtgta tgaacatacc attaattgtt ccttctctag ttgataatta tcgaaaaata
      601 aatatagcaa ataataaatc aaacaacgat ttaaccaaaa gagaaaaaga atgtttagcg
      661 tgggcatgcg aaggaaaaag ctcttgggat atttcaaaaa tattaggttg cagtgagcgt
      721 actgtcactt tccatttaac caatgcgcaa atgaaactca atacaacaaa ccgctgccaa
      781 agtatttcta aagcaatttt aacaggagca attgattgcc catactttaa aaattaataa
      841 cactgatagt gctagtgtag atcactacta gagccaggca tcaaataaaa cgaaaggctc
      901 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
      961 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagag acctgtagga
     1021 tcgtacaggt ttacgcaaga aaatggtttg ttatagtcga ataaatacta gagtcacaca
     1081 ggaaagtact agatgcgtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
     1141 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
     1201 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
     1261 tggccaacac ttgtcactac tttcggttat ggtgttcaat gctttgcgag atacccagat
     1321 catatgaaac agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga
     1381 actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt
     1441 gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt
     1501 cttggacaca aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa
     1561 caaaagaatg gaatcaaagt taacttcaaa attagacaca acattgaaga tggaagcgtt
     1621 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca
     1681 gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac
     1741 cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta
     1801 tacaaataat actagtagcg gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc
     1861 ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac
     1921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccaca ggctccgccc
     2101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3541 gatcttaccg ctgttgagat ccagttcgat ataacccact cgtgcaccca actgatcttc
     3601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3841 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt
     3901 cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct
     3961 agag
//
