LOCUS       40924_38773        7413 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSB4K5-CadC-VHH-Caffeine, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7413)
  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 7413)
  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 7413)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7413)
  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..7413
                     /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..1190)
                     /label=PcadBA
                     /note="PcadBA"
                     /regulatory_class="promoter"
     terminator      1238..1265
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    1368..1384
                     /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    1391..1407
                     /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      1427..1450
                     /label=RBS benchmarking
                     /note="RBS benchmarking"
                     /regulatory_class="ribosome_binding_site"
     CDS             2393..2422
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     terminator      2520..2591
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2607..2634
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2875..3954
                     /label=lacI
                     /note="lac repressor"
     terminator      4073..4104
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     CDS             complement(4206..5018)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(5379..6326)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(6374..6596)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(7088..7182)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      7219..7278
                     /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(7338..7367)
                     /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 aaactggcga taagattact cacgaaaaaa ggattaatcc taaagattag
     1081 gtgaaataac acaaaagttt ctgtaagtga gaacttgagg ttttttatta acacatcagg
     1141 atcgcaagtt gatatcatga aaagataaac atttaatgtt tacaatggat aaggaacggt
     1201 tatttctgcg tagatctatc ttacacagca tcacactggc tcaccttcgg gtgggccttt
     1261 ctgcgtttat atactagaga gagaatataa aaagccagat tattaatccg gcttttttat
     1321 tatttaggca actgaaacga ttcggatcct gtattactat tcttaaattg tgagcggata
     1381 acaattgaca ttgtgagcgg ataacaagat actgagcaca gctagcgatt aactaataag
     1441 gaggacaaac atgcaacaac ctgtagttcg cgttggcgaa tggcttgtta ctccgtccat
     1501 aaaccaaatt agccgcaatg ggcgtcaact tacccttgag ccgagattaa tcgatcttct
     1561 ggttttcttt gctcaacaca gtggcgaagt acttagcagg gatgaactta tcgataatgt
     1621 ctggaagaga agtattgtca ccaatcacgt tgtgacgcag agtatctcag aactacgtaa
     1681 gtcattaaaa gataatgatg aagatagtcc tgtctatatc gctactgtac caaagcgcgg
     1741 ctataaatta atggtgccgg ttatctggta cagcgaagaa gagggagagg aaataatgct
     1801 atcttcgcct ccccctatac cagaggcggt tcctgccaca gattctccct cccacagtct
     1861 taacattcaa aacaccgcaa cgccacctga acaatcccca gttaaaagca aacgaggtgg
     1921 cccaggtctg ttgctgttgc tgttgctgtt gctgttgctg ttgctgttgc tgttgggtcc
     1981 aggtggcgaa gttcaactgc aagcttctgg cggcggcctg gttcaagctg gcggctctct
     2041 gcgtctgtct tgcactgctt ctggccgtac tggcactatc tactctatgg cttggttccg
     2101 tcaagctcct ggcaaagaac gtgaattcct ggctactgtt ggctggtctt ctggcatcac
     2161 ttactacatg gattctgtta aaggccgttt cactatctct cgtgataacg ctaaaaactc
     2221 tgcttacctg caaatgaact ctctgaaacc tgaagatact gctgtttact actgcactgc
     2281 tactcgtgct tactctgttg gctacgatta ctggggccaa ggcactcaag ttactgtttc
     2341 tcatgctgct gctggcgctc ctgttcctta ccctgatcct ctggaacctc gtgaacaaaa
     2401 actgatctct gaagaagatc tgtaaacttt atctgagaat agtcaatctt cggaaatccc
     2461 aggtggcatg ctaaaagtct cgtaaagcgt tctatcaata acccgttggt gccaggcatc
     2521 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     2581 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
     2641 ccgtctcaga atcggccgtg aacaataaaa tagtttcggt ttgcatttaa atcttacata
     2701 tgtaatactt tcaaagacta catttgtaag atttgatgtt tgagtcggct gaaagatcgt
     2761 acgtaccaat tattgtttcg tgattgttca agccataaca ctgtagggat agtggaaaga
     2821 gtgcttcatc tggttacgat caatcaaata ttcaaacgga gggagacgat tttgatgaaa
     2881 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     2941 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     3001 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     3061 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     3121 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     3181 ggcgtcgaag cctgtaaaac ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     3241 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     3301 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     3361 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     3421 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     3481 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     3541 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     3601 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     3661 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     3721 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     3781 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     3841 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3901 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgataa
     3961 taaaaggtcc cgtctgaact tactgtgaat tcgactaatt attgaccact tccgagtaga
     4021 atcgtgcttc agtaagagag tcactaaggg ttagttagtt agattagcag aaagtcaaaa
     4081 gcctccgacc ggaggctttt gactaaaact tcccttgggg ttatcattgg ggctcactca
     4141 aaggcggtaa tcagataaaa aaaatcctta gctttcgcta aggatgattt ctgctagtat
     4201 tattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     4261 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     4321 caaagaatgg caaggtcctg gtaacggtct gcgattccga cccgtccaac atcaatacaa
     4381 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     4441 actgaatccg gtgagaatgg caagagcttg tgcatttctt tccagacttg ttcaacaggc
     4501 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt catgcgtgat
     4561 tgcgcctgag caagacgaaa tacacgatcg ctgttaaaag gacaattaca aacaggaatc
     4621 gaatgtaacc ggcgcaggaa cacggccagc gcatcaacaa tattttcacc tgaatcagga
     4681 tattcttcta atacctggaa ggctgttttc ccaggaatcg cggtggtgag taaccacgca
     4741 tcatcaggag tacggataaa atgcttgatg gtcgggagag gcataaactc cgtcagccag
     4801 ttgagacgga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     4861 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
     4921 acattatcgc gagcccattt atacccatat aaatcagcgt ccatgttgga gtttaagcgc
     4981 ggacgggagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     5041 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     5101 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     5161 gatcagctct agtagttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa
     5221 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg
     5281 acagttttcc ctttgatatc taacggtgaa cagttgttct acttttgttt gttagtcttg
     5341 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt
     5401 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca
     5461 tgcttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca
     5521 gttaaagcat cgtgtagtgt ttttcttagt ccgttacgta ggtaggaatc tgatgtaatg
     5581 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga
     5641 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta
     5701 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc
     5761 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat
     5821 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat
     5881 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta
     5941 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat
     6001 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta
     6061 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat
     6121 acaccataag cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac
     6181 gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag
     6241 cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt
     6301 gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta
     6361 taccaattga gatgggctag tcaatgataa ttacatgtcc ttttcctttg agttgtgggt
     6421 atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta
     6481 aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga
     6541 ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta
     6601 ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa
     6661 aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg
     6721 aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca
     6781 gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt
     6841 atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg
     6901 gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct
     6961 gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg
     7021 ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag
     7081 tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     7141 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgtaaact
     7201 tggtctgaca gctctagctc cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt
     7261 tctttaaaac cgaaaagatt acttcgcgtt tgccacctga cgtctaagaa aaggaatatt
     7321 cagcaatttg cccgtgccga agaaaggccc acccgtgaag gtgagccagt gagttgattg
     7381 ctacgtaatt agttagttag cccttagtga ctc
//
