LOCUS       Exported                7408 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSCT32, complete sequence.
ACCESSION   KY078890
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7408)
  AUTHORS   Chen S.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2016) School of Life Sciences, Qingdao 
            Agricultural University, 700 Changcheng Road, Qingdao, Shandong 
            266109, PR China
REFERENCE   2  (bases 1 to 7408)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7408
                     /organism="Cloning vector pSCT32"
                     /lab_host="Gram negative bacterium"
                     /mol_type="other DNA"
                     /label=derived from Escherichia coli DH5a sequence
                     /note="derived from Escherichia coli DH5a sequence"
                     /db_xref="taxon:1983414"
     promoter        complement(1..103)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      404..1023
                     /label=from pBR322
                     /note="from pBR322"
     rep_origin      420..1008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(1256..1684)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      1361..1667
                     /label=from f1
                     /note="from f1"
     promoter        1982..2059
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2060..3142
                     /codon_start=1
                     /product="laqI"
                     /label=laqI
                     /protein_id="ARN18287.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     regulatory      3374..3431
                     /regulatory_class="promoter"
                     /note="Trc"
     protein_bind    3411..3427
                     /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)."
     CDS             3450..4133
                     /codon_start=1
                     /product="mRFP"
                     /label=mRFP
                     /protein_id="ARN18288.1"
                     /translation="MDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVT
                     KGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVT
                     QDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMRLKL
                     KDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHSTGAH
                     HHHHH"
     CDS             4113..4130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      4233..4390
                     /regulatory_class="terminator"
                     /note="rrnB"
     terminator      4353..4439
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      4356..4399
                     /regulatory_class="terminator"
                     /note="rrnB_T1"
     regulatory      4531..4558
                     /regulatory_class="terminator"
                     /note="rrnB_T2"
     terminator      4531..4558
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(4703..5299)
                     /codon_start=1
                     /product="TraI"
                     /label=TraI
                     /protein_id="ARN18289.1"
                     /translation="MRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANTLPAVMAE
                     VMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQRVSAVHHD
                     TDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQRVSENRA
                     NDMERHAGVESLVGWIIYSASQSLIKTVSASLIQM"
     CDS             complement(5355..5726)
                     /codon_start=1
                     /product="TraJ"
                     /label=TraJ
                     /protein_id="ARN18290.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5759..5868
                     /note="incP origin of transfer"
     oriT            5806..5904
                     /label=from RP4
                     /note="from RP4"
     CDS             complement(6749..7408)
                     /codon_start=1
                     /product="CAT"
                     /label=CAT
                     /protein_id="ARN18291.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"