LOCUS       Exported                5090 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHTSUB-105, complete sequence.
ACCESSION   AY902486
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5090)
  AUTHORS   Basu S, Gerchman Y, Collins CH, Arnold FH, Weiss R.
  TITLE     A synthetic multicellular system for programmed pattern formation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5090)
  AUTHORS   Basu S, Gerchman Y, Collins CH, Arnold FH, Weiss R.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JAN-2005) Electrical Engineering, Princeton 
            University, J-319, Engineering Quadrangle, Olden Street, Princeton, 
            NJ 08544, USA
REFERENCE   3  (bases 1 to 5090)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5090
                     /organism="Expression vector pHTSUB-105"
                     /mol_type="other DNA"
                     /label=high sensitive high threshold plasmid
                     /note="high sensitive high threshold plasmid"
                     /db_xref="taxon:313407"
     CDS             complement(61..1143)
                     /codon_start=1
                     /gene="LacI-M1"
                     /product="LacI-M1"
                     /label=LacI-M1
                     /note="codon optimized LacI"
                     /protein_id="AAX07427.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(61..1143)
                     /gene="LacI-M1"
                     /label=LacI-M1
     regulatory      1017..1026
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     RBS             1147..1155
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             1371..2123
                     /codon_start=1
                     /product="LuxR-mut"
                     /label=LuxR-mut
                     /note="R-protein from Vibrio fischeri mutated for better 
                     sensitivity"
                     /protein_id="AAX07428.1"
                     /translation="MKNINADDTYRIINKIKACRSNNDINQCLSDMTKMVHCEYYLLAI
                     IYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYSNSNHSPINWNIFENNAVNK
                     KSPNVIKEAKTSGLITGFSFPIHTANNGFGMLSFAHSEKDNYIDSLFLHACMNIPLIVP
                     SLVDNYRKINIANNKSNNDLTKREKECLAWACEGKSSWDISKILGCSERTVTFHLTNAQ
                     MKLNTTNRCQSISKAILTGAIDCPYFKN"
     terminator      2191..2277
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2374..2395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2410..2440
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2448..2464
                     /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)."
     primer_bind     2472..2488
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             2553..2564
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2571..3326
                     /codon_start=1
                     /product="GFP-LVA"
                     /label=GFP-LVA
                     /note="green flourescent protein with LVA fast decay tag"
                     /protein_id="AAX07429.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKRPAANDENYALVA"
     CDS             2571..3284
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     rep_origin      complement(3443..4031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4119..4213
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4234..4893)
                     /codon_start=1
                     /gene="Cm(r)"
                     /product="chloromphenicol resistance protein"
                     /label=Cm(r)
                     /protein_id="AAX07430.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(4234..4893)
                     /gene="Cm(r)"
                     /label=Cm(r)
     promoter        complement(4894..4996)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"