pACYCDuet-1 Plasmid
PVT0102 2ug
pACYCDuet-1 plasmid information
Promoter: T7/Lac
Replicon: p15A ori
Terminator: T7 Terminator
Plasmid classification: large intestine series plasmid; other large intestine plasmid; large intestine double frame plasmid
Plasmid size: 4008bp
Plasmid Tags: N-6 x His, C-S
Prokaryotic resistance: chloramphenicol Chl (50 g/ml)
Clone strain: DH5 alpha and other Escherichia coli
Culture conditions: 37, aerobic, LB
Expression host: BL21 (DE3) and other Escherichia coli
Culture conditions: 37, aerobic, LB
Induction: IPTG or lactose and its analogues
Primers for 5'sequencing: ACYCDuetUP1 (GGATCTCGACGCTCTCCCT) DuetUP2 (TTGTACACGGCCGCATAATC)
pACYCDuet-1 plasmid Description
pACYCDuet-1 plasmid is designed for the coexpression of two target genes. The vector contains two multiple cloning sites (MCS), each of which is preceded by a T7 promoter/lac operator and ribosome binding site (rbs). The vector also carries the P15A replicon, lacI gene and chloramphenicol resistance gene. This vector can be used in combination with pETDuet™-1 in an appropriate host strain for the coexpression of up to 4 target genes. Genes inserted into MCS1 can be sequenced using the ACYCDuetUP1 Primer and DuetDOWN1 Primer . Genes inserted into MCS2 can be sequenced using the DuetUP2 Primer and T7 Terminator Primer .
pACYCDuet-1 plasmid Sequence
LOCUS Exported 4008 bp ds-DNA circular SYN 27-8-2015
DEFINITION synthetic circular DNA
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4008)
TITLE Direct Submission
JOURNAL Exported 2015-8-27
FEATURES Location/Qualifiers
source 1..4008
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 3..27
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
CDS 83..100
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
promoter 214..232
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 233..257
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
CDS 366..410
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
terminator 462..509
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(729..1388)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(1389..1491)
/note="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin complement(2017..2561)
/direction=LEFT
/note="p15A ori"
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
CDS complement(2786..3868)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="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
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(3869..3946)
/gene="lacI"
/note="lacI promoter"
promoter 3992..2
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
481 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
541 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
601 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
661 ccgcttatta tcacttattc aggcgtagca ccaggcgttt aagggcacca ataactgcct
721 taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
781 ctgccgacat ggaagccatc acagacggca tgatgaacct gaatcgccag cggcatcagc
841 accttgtcgc cttgcgtata atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc
901 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
961 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
1021 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
1081 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
1141 accagctcac cgtctttcat tgccatacgg aactccggat gagcattcat caggcgggca
1201 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
1261 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
1321 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
1381 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
1441 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
1501 tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat
1561 tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct
1621 gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc
1681 tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc
1741 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
1801 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
1861 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
1921 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
1981 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
2041 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
2101 ataccaggcg tttcccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
2161 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
2221 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
2281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
2341 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
2401 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
2461 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
2521 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
2581 agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
2641 gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga
2701 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
2761 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
2821 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
2881 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
2941 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
3001 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
3061 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
3121 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
3181 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
3241 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
3301 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
3361 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
3421 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
3481 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
3541 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
3601 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
3661 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
3721 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
3781 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
3841 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
3901 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
3961 tctcccttat gcgactcctg cattaggaaa ttaatacgac tcactata
//
Caution:
Product is for research use only!
Search name
pACYCDuet-1,Plasmid pACYCDuet-1,pACYCDuet-1 vector
