LOCUS       sequence-251038-        5999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the synthase LuxI for the quorum-sensing molecule C6-HSL..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5999)
  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 5999)
  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: Sender plasmid
FEATURES             Location/Qualifiers
     source          1..5999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             651..683
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     misc_feature    715..735
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     promoter        813..831
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    832..856
                     /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)."
     CDS             965..1009
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1047..1065)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1061..1108
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1145..1600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1232..1251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1442..1463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1715..2506,2507..2575))
                     /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     2338..2357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2576..2668)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2749..3337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3238..3257
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3491..3508
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3523..3662
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3588..3607)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3748..3770
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3764..3955)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4527..5609)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5571..5590
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5610..5687)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5893..5912
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gagaaagagg
       61 ggaaatacta gatgactata atgataaaaa aatcggattt tttggcaatt ccatcggagg
      121 agtataaagg tattctaagt cttcgttatc aagtgtttaa gcaaagactt gagtgggact
      181 tagttgtaga aaataacctt gaatcagatg agtatgataa ctcaaatgca gaatatattt
      241 atgcttgtga tgatactgaa aatgtaagtg gatgctggcg tttattacct acaacaggtg
      301 attatatgct gaaaagtgtt tttcctgaat tgcttggtca acagagtgct cccaaagatc
      361 ctaatatagt cgaattaagt cgttttgctg taggtaaaaa tagctcaaag ataaataact
      421 ctgctagtga aattacaatg aaactatttg aagctatata taaacacgct gttagtcaag
      481 gtattacaga atatgtaaca gtaacatcaa cagcaataga gcgattttta aagcgtatta
      541 aagttccttg tcatcgtatt ggagacaaag aaattcatgt attaggtgat actaaatcgg
      601 ttgtattgtc tatgcctatt aatgaacagt ttaaaaaagc agtcttaaat gctgcaaacg
      661 acgaaaacta cgctttagta gcttaataac tctgatagtg ctagtgtaga tctctactag
      721 tagcggccgc tgcaggtcga caagcttgcg gccgcataat gcttaagtcg aacagaaagt
      781 aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag gggaattgtg
      841 agcggataac aattccccat cttagtatat tagttaagta taagaaggag atatacatat
      901 ggcagatctc aattggatat cggccggcca cgcgatcgct gacgtcggta ccctcgagtc
      961 tggtaaagaa accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc
     1021 agcttaatta acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1081 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     1141 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1201 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1261 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1321 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1381 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1441 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1501 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1561 tttaacgcga attttaacaa aatattaacg tttacaattt ctggcggcac gatggcatga
     1621 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1681 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1741 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1801 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1861 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1921 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1981 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2041 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2101 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2161 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2221 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     2281 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     2341 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2401 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2461 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2521 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2581 tcctttttca atcatgattg aagcatttat cagggttatt gtctcatgag cggatacata
     2641 tttgaatgta tttagaaaaa taaacaaata ggtcatgacc aaaatccctt aacgtgagtt
     2701 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2761 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2821 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2881 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2941 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3001 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3061 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3121 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3181 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3241 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3301 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3361 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3421 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3481 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     3541 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     3601 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3661 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3721 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3781 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3841 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3901 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3961 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     4021 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     4081 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     4141 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     4201 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     4261 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     4321 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     4381 gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc caccggaagg
     4441 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     4501 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4561 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     4621 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     4681 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     4741 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     4801 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     4861 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     4921 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     4981 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     5041 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     5101 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     5161 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     5221 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     5281 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     5341 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     5401 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     5461 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     5521 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     5581 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     5641 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     5701 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     5761 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     5821 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     5881 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     5941 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaat
//
