LOCUS       Exported                5157 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHTSUB-106, complete sequence.
ACCESSION   AY952933
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5157)
  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  JOURNAL   Unpublished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 5157)
  AUTHORS   Basu S, Gerchman Y, Collins CH, Arnold FH, Weiss R.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JAN-2005) Electrical Engineering, Princeton 
            University, B-312, E-Quad, Olden Street, Princeton, NJ 08544, USA
REFERENCE   3  (bases 1 to 5157)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5157
                     /organism="Cloning vector pHTSUB-106"
                     /mol_type="other DNA"
                     /label=low copy plasmid
                     /note="low copy plasmid"
                     /db_xref="taxon:316972"
     CDS             complement(122..1204)
                     /codon_start=1
                     /gene="lacIM1"
                     /product="LacI-M1"
                     /label=lacIM1
                     /note="codon-optimized LacI"
                     /protein_id="AAX29977.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(122..1204)
                     /gene="lacIM1"
                     /label=lacIM1
     regulatory      1078..1087
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     RBS             1208..1216
                     /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             1428..2180
                     /codon_start=1
                     /gene="luxR"
                     /product="LuxR"
                     /label=luxR
                     /note="R-protein from Vibrio fischeri"
                     /protein_id="AAX29978.1"
                     /translation="MKNINADDTYRIINKIKACRSNNDINQCLSDMTKMVHCEYYLLAI
                     IYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYSNSNHSPINWNIFENNAVNK
                     KSPNVIKEAKTSGLITGFSFPIHTANNGFGMLSFAHSEKDNYIDSLFLHACMNIPLIVP
                     SLVDNYRKINIANNKSNNDLTKREKECLAWACEGKSSWDISKILGCSERTVTFHLTNAQ
                     MKLNTTNRCQSISKAILTGAIDCPYFKN"
     gene            1428..2180
                     /gene="luxR"
                     /label=luxR
     terminator      2291..2377
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2502..2523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2538..2568
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2576..2592
                     /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     2600..2616
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             2681..2692
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2699..3454
                     /codon_start=1
                     /gene="gfpLVA"
                     /product="green fluorescent protein LVA"
                     /label=gfpLVA
                     /note="GFP-LVA; green flourescent protein with LVA fast 
                     decay tag"
                     /protein_id="AAX29979.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKRPAANDENYALVA"
     gene            2699..3454
                     /gene="gfpLVA"
                     /label=gfpLVA
     CDS             2699..3412
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     rep_origin      complement(3571..4159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4247..4341
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4362..5021)
                     /codon_start=1
                     /gene="cm(r)"
                     /product="chloroamphenicol resistance protein"
                     /label=cm(r)
                     /note="Cm(r)"
                     /protein_id="AAX29980.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(4362..5021)
                     /gene="cm(r)"
                     /label=cm(r)
     promoter        complement(5022..5124)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"