LOCUS       40924_38783        7056 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSB4K5-LexA-VHH-Caffeine, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Chang HJ, Mayonove P, Zavala A, De Visch A, Minard P, Cohen-Gonsaud 
            M, Jerome B.
  TITLE     A modular receptor platform to expand the sensing repertoire of 
            bacteria
  JOURNAL   ACS Synth Biol (2017) In press
  PUBMED    28946740
REFERENCE   2  (bases 1 to 7056)
  AUTHORS   Chang H-J., Mayonove P, De Visch A, Minard P, Cohen-Gonsaud M, 
            Bonnet J.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-OCT-2017) Biophysics, Centre de Biochimie Structurale 
            de Montpellier, 29 Rue de Navacelles, Montpellier 34090, France
REFERENCE   3  (bases 1 to 7056)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7056)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-OCT-2017) Biophysics, Centre de Biochimie Structurale de 
            Montpellier, 29 Rue de Navacelles, Montpellier 34090, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      complement(89..253)
                     /label=Terminator Benchmarking
                     /note="Terminator Benchmarking"
                     /regulatory_class="terminator"
     CDS             complement(297..1010)
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     regulatory      complement(1011..1022)
                     /label=RBS K143021
                     /note="RBS K143021"
                     /regulatory_class="ribosome_binding_site"
     regulatory      complement(1031..1111)
                     /label=PsulA408
                     /note="PsulA408"
                     /regulatory_class="promoter"
     terminator      1157..1184
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    1287..1303
                     /label=lac operator
                     /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)."
     protein_bind    1310..1326
                     /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)."
     regulatory      1346..1369
                     /label=RBS benchmarking
                     /note="RBS benchmarking"
                     /regulatory_class="ribosome_binding_site"
     CDS             1370..1621
                     /label=LexA BD
                     /note="DNA-binding domain of E. coli LexA"
     CDS             2036..2065
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     terminator      2163..2234
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2250..2277
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2518..3597
                     /label=lacI
                     /note="lac repressor"
     terminator      3716..3747
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     CDS             complement(3849..4661)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(5022..5969)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(6017..6239)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(6731..6825)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      6862..6921
                     /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."
     terminator      complement(6981..7010)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ctcggatacc cttactctgt tgaaaacgaa tagataggtt tgctcgtagt ttaccacgga
       61 tacagacagt gataatctta ggcgcgccga atcttgcttg gcaaggttct tttttttttt
      121 caaaaaagca cgaaaaaagc acccataagg taaacttttg agtgctttta aatacgcttc
      181 acagtttctt cttcattttt aacaaaaaac ccagctcatt gagctgggtt taagctggct
      241 tggcggcgtc ctaagattac tactgataac cactgttgat tgggataccc gtattattta
      301 taaagttcgt ccataccgtg agtgatacct gctgctgtaa caaattcaag aagtaccata
      361 tggtcacgtt tttcgttagg atcttttgaa agaactgatt gagtagaaag gtagtggtta
      421 tctggaagca atactggtcc gtctccgatt ggtgtatttt gttgataatg atcagccaat
      481 tgaacagatc catcttcaac attgtgacga attttaaaat tagctttaat accgtttttt
      541 tgtttgtcag ctgtgatata aacattatga gagttgaagt tgtattcaag tttgtgtccc
      601 aaaatgttac catcttcttt gaaatcgatt cctttcaatt cgatacgatt aacaagagtg
      661 tcaccttcga attttacttc agcacgtgtt ttatatgttc cgtcatcttt aaatgaaatt
      721 gtacgttctt gtacataacc ttctggcata gcagatttaa aaaaatcgtg acgtttcata
      781 tgatcaggat aacgtgaaaa acattgtact ccatatgtca atgttgtaac aagagtaggc
      841 caaggaacag gaagttttcc agttgtacaa atgaatttaa gtgtcaattt accgttagtt
      901 gcatcacctt caccttcacc acgtacagaa aatttgtgac cgttaacatc accatccaat
      961 tcaaccaaga taggtactac acctgtaaaa agttcttctc cttttgacat ttcaccacct
     1021 ttactagtcc agcccctgtg agttaccgta tggatgtacg gtacatccag tgacaacaaa
     1081 gatcaaccct attttcggaa agagcctcga aggaacggtt atttctgcgt agatctatct
     1141 tacacagcat cacactggct caccttcggg tgggcctttc tgcgtttata tactagagag
     1201 agaatataaa aagccagatt attaatccgg cttttttatt atttaggcaa ctgaaacgat
     1261 tcggatcctg tattactatt cttaaattgt gagcggataa caattgacat tgtgagcgga
     1321 taacaagata ctgagcacag ctagcgatta actaataagg aggacaaaca tgaaagcgct
     1381 gaccgcgcgt caacaagaag ttttcgacct gatccgtgac cacatcagcc agacgggtat
     1441 gccacctacc cgtgcggaaa tcgcgcaacg tctgggcttc cgtagcgcaa gctctgctga
     1501 agagcacctg aaagcgctgg cccgtaaggg tgttatcgaa atcgtttccg gtgcgagccg
     1561 cggtatccgt ctgctgcagg aagaagaaga aggtctgccg ctggtgggcc gtgtggctgc
     1621 cggcgaacca gaagttcaac tgcaagcttc tggcggcggc ctggttcaag ctggcggctc
     1681 tctgcgtctg tcttgcactg cttctggccg tactggcact atctactcta tggcttggtt
     1741 ccgtcaagct cctggcaaag aacgtgaatt cctggctact gttggctggt cttctggcat
     1801 cacttactac atggattctg ttaaaggccg tttcactatc tctcgtgata acgctaaaaa
     1861 ctctgcttac ctgcaaatga actctctgaa acctgaagat actgctgttt actactgcac
     1921 tgctactcgt gcttactctg ttggctacga ttactggggc caaggcactc aagttactgt
     1981 ttctcatgct gctgctggcg ctcctgttcc ttaccctgat cctctggaac ctcgtgaaca
     2041 aaaactgatc tctgaagaag atctgtaaac tttatctgag aatagtcaat cttcggaaat
     2101 cccaggtggc atgctaaaag tctcgtaaag cgttctatca ataacccgtt ggtgccaggc
     2161 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     2221 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     2281 ataccgtctc agaatcggcc gtgaacaata aaatagtttc ggtttgcatt taaatcttac
     2341 atatgtaata ctttcaaaga ctacatttgt aagatttgat gtttgagtcg gctgaaagat
     2401 cgtacgtacc aattattgtt tcgtgattgt tcaagccata acactgtagg gatagtggaa
     2461 agagtgcttc atctggttac gatcaatcaa atattcaaac ggagggagac gattttgatg
     2521 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     2581 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     2641 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
     2701 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
     2761 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
     2821 agcggcgtcg aagcctgtaa aacggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
     2881 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
     2941 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc
     3001 tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa
     3061 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg
     3121 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
     3181 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
     3241 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
     3301 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
     3361 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
     3421 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
     3481 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
     3541 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     3601 taataaaagg tcccgtctga acttactgtg aattcgacta attattgacc acttccgagt
     3661 agaatcgtgc ttcagtaaga gagtcactaa gggttagtta gttagattag cagaaagtca
     3721 aaagcctccg accggaggct tttgactaaa acttcccttg gggttatcat tggggctcac
     3781 tcaaaggcgg taatcagata aaaaaaatcc ttagctttcg ctaaggatga tttctgctag
     3841 tattattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     3901 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     3961 ttccaaagaa tggcaaggtc ctggtaacgg tctgcgattc cgacccgtcc aacatcaata
     4021 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     4081 acgactgaat ccggtgagaa tggcaagagc ttgtgcattt ctttccagac ttgttcaaca
     4141 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcatgcgt
     4201 gattgcgcct gagcaagacg aaatacacga tcgctgttaa aaggacaatt acaaacagga
     4261 atcgaatgta accggcgcag gaacacggcc agcgcatcaa caatattttc acctgaatca
     4321 ggatattctt ctaatacctg gaaggctgtt ttcccaggaa tcgcggtggt gagtaaccac
     4381 gcatcatcag gagtacggat aaaatgcttg atggtcggga gaggcataaa ctccgtcagc
     4441 cagttgagac ggaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     4501 agaaacaact ctggcgcatc gggcttccca tacaagcgat agattgtcgc acctgattgc
     4561 ccgacattat cgcgagccca tttataccca tataaatcag cgtccatgtt ggagtttaag
     4621 cgcggacggg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     4681 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     4741 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     4801 aaggatcagc tctagtagtt acattgtcga tctgttcatg gtgaacagct ttaaatgcac
     4861 caaaaactcg taaaagctct gatgtatcta tcttttttac accgttttca tctgtgcata
     4921 tggacagttt tccctttgat atctaacggt gaacagttgt tctacttttg tttgttagtc
     4981 ttgatgcttc actgatagat acaagagcca taagaacctc agatccttcc gtatttagcc
     5041 agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga
     5101 tcatgcttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt
     5161 gcagttaaag catcgtgtag tgtttttctt agtccgttac gtaggtagga atctgatgta
     5221 atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg
     5281 agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc
     5341 ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa
     5401 acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta
     5461 aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt
     5521 aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga
     5581 ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact
     5641 aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct
     5701 ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct
     5761 aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg
     5821 aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca
     5881 cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca
     5941 tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca
     6001 ctataccaat tgagatgggc tagtcaatga taattacatg tccttttcct ttgagttgtg
     6061 ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct
     6121 gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat
     6181 agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca
     6241 ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta
     6301 caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg
     6361 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
     6421 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
     6481 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
     6541 agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
     6601 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
     6661 tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtccc
     6721 tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     6781 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgtaa
     6841 acttggtctg acagctctag ctccggcaaa aaaacgggca aggtgtcacc accctgccct
     6901 ttttctttaa aaccgaaaag attacttcgc gtttgccacc tgacgtctaa gaaaaggaat
     6961 attcagcaat ttgcccgtgc cgaagaaagg cccacccgtg aaggtgagcc agtgagttga
     7021 ttgctacgta attagttagt tagcccttag tgactc
//
