LOCUS       Exported                7413 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSB4K5-CadC-VHH-Caffeine, complete sequence.
ACCESSION   MG210478
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7413
                     /organism="Cloning vector pSB4K5-CadC-VHH-Caffeine"
                     /mol_type="other DNA"
                     /db_xref="taxon:2049006"
     regulatory      complement(89..253)
                     /regulatory_class="terminator"
                     /note="Terminator Benchmarking"
     CDS             complement(294..1010)
                     /codon_start=1
                     /product="sfGFP"
                     /label=sfGFP
                     /note="super folder green fluorescent protein"
                     /protein_id="ATO93722.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      complement(1011..1022)
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS K143021"
     regulatory      complement(1031..1190)
                     /regulatory_class="promoter"
                     /note="PcadBA"
     regulatory      1231..1325
                     /regulatory_class="terminator"
                     /note="B014"
     terminator      1238..1265
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      1366..1420
                     /regulatory_class="promoter"
                     /note="pLacO1"
     protein_bind    1368..1384
                     /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)."
     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
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS benchmarking"
     CDS             1451..2425
                     /codon_start=1
                     /product="CadC-VHH-Caffeine"
                     /label=CadC-VHH-Caffeine
                     /note="contains CadC DNA binding domain fused with 
                     VHH-Caffeine"
                     /protein_id="ATO93723.1"
                     /translation="MQQPVVRVGEWLVTPSINQISRNGRQLTLEPRLIDLLVFFAQHSG
                     EVLSRDELIDNVWKRSIVTNHVVTQSISELRKSLKDNDEDSPVYIATVPKRGYKLMVPV
                     IWYSEEEGEEIMLSSPPPIPEAVPATDSPSHSLNIQNTATPPEQSPVKSKRGGPGLLLL
                     LLLLLLLLLLLLGPGGEVQLQASGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQAPGK
                     EREFLATVGWSSGITYYMDSVKGRFTISRDNAKNSAYLQMNSLKPEDTAVYYCTATRAY
                     SVGYDYWGQGTQVTVSHAAAGAPVPYPDPLEPREQKLISEEDL"
     CDS             2393..2422
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2512..2680
                     /regulatory_class="terminator"
                     /note="B0015"
     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..3957
                     /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
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     terminator      4073..4104
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4203..5018)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5376..6326)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      6374..6596
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      7088..7182
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      7338..7367
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"