LOCUS pACYC-T7 5937 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5937)
AUTHORS Wayne N, Bolon DN
TITLE Dimerization of Hsp90 is required for in vivo function. Design and
analysis of monomers and dimers.
JOURNAL J Biol Chem. 2007 Nov 30;282(48):35386-95. Epub 2007 Oct 1.
PUBMED 17908693
REFERENCE 2 (bases 1 to 5937)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem."; date: "2007-11-30"; volume: "282(48)"; pages: "35386-95.
Epub 2007 Oct 1"
FEATURES Location/Qualifiers
source 1..5937
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 6..25
/label=T7
/note="T7 promoter, forward primer"
promoter 6..24
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 25..49
/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 105..122
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 2315..2359
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(2397..2415)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2411..2458
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(2679..3338)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 3253..3272
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(3339..3441)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(3833..3850)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3967..4512)
/direction=LEFT
/label=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.
"
primer_bind complement(4047..4066)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4737..5819)
/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
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 5781..5800
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5820..5897)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctgtagaaat
61 aattttgttt aactttaata aggagatata ccatgggcag cagccatcat catcatcatc
121 attctagtca tatggctagt gaaacttttg aatttcaagc tgaaattact cagttgatga
181 gtttgatcat caacaccgtc tattctaaca aggaaatttt cttgagagaa ctgatatcta
241 atgcctcgga tgcgttggat aaaattagat acaaatcttt gtctgatcca aagcaattgg
301 aaacagaacc agatctcttt attagaatca ctccaaagcc agagcaaaaa gttttggaaa
361 tcagagattc tggtattggt atgaccaagg ctgaattgat taataacttg ggtaccattg
421 ccaagtctgg taccaaagcc ttcatggaag ctctatctgc tggtgccgat gtatccatga
481 ttggtcaatt cggtgttggt ttttactctt tattcttagt tgccgacaga gttcaggtta
541 tttcaaagag caacgacgac gaacaataca tctgggaatc caacgctggt ggttctttca
601 ctgttactct agacgaagtt aatgaaagaa ttggtagggg taccatcttg aggttattct
661 tgaaagatga ccaattggag tacttggaag aaaagagaat aaaggaagtt atcaagagac
721 attctgagtt cgtggcctac ccaatccaat tagtcgtcac caaggaagtt gaaaaggaag
781 ttccaattcc agaagaagaa aagaaagacg aggaaaagaa ggatgaggaa aagaaggatg
841 aagacgacaa gaaaccaaaa ctcgaggaag tcgatgaaga agaggaaaag aagccaaaga
901 cgaaaaaagt taaagaagaa gttcaagaga tagaagaact aaacaagact aagcctttgt
961 ggactagaaa cccatctgat atcactcaag aagaatacaa tgctttctat aagtctattt
1021 caaacgactg ggaagaccca ttgtacgtta agcatttctc cgttgaaggt caattggaat
1081 ttagagctat cttattcatt ccaaagagag caccattcga cttgtttgag agtaaaaaga
1141 agaagaataa tatcaagttg tacgttcgtc gtgttttcat cactgatgaa gctgaagact
1201 tgattccaga gtggttatct ttcgtcaagg gtgttgttga ctctgaggat ttaccattga
1261 atttgtccag agaaatgtta caacaaaata agatcatgaa ggttattaga aagaacattg
1321 tcaaaaagtt gattgaagcc ttcaacgaaa ttgctgaaga ctctgaacaa tttgaaaagt
1381 tctactcggc tttctccaaa aatatcaagt tgggtgtaca tgaagatacc caaaacaggg
1441 ctgctttggc taagttgtta cgttacaact ctaccaagtc cgtagatgag ttgacttcct
1501 taactgatta cgttaccaga atgccagaac accaaaagaa catctactac atcactggtg
1561 aatctctaaa ggctgtcgaa aagtctccat ttttggatgc cttgaaggct aaaaacttcg
1621 aggttttgtt cttgaccgac ccaattgatg aatacgcctt cactcaattg aaggaattcg
1681 aaggtaaaac tttggttgac attactaaag atttcgaatt ggaagaaact gacgaagaaa
1741 aagctgaaag agagaaggag atcaaagaat atgaaccatt gaccaaagct ttgaaagaaa
1801 ttttgggtga ccaagtggag aaagttgttg tttcttacaa attgttggat gccccagctg
1861 ctatcagaac tggtcaattt ggttggtctg ctaacatgga aagaatcatg aaggctcaag
1921 ccttgagaga ctcttccatg tcctcctaca tgtcttccaa gaagactttc gaaatttctc
1981 caaaatctcc aattatcaag gaattgaaaa agagagttga cgaaggtggt gctcaagaca
2041 agactgtcaa ggacttgact aagttattat atgaaactgc tttgttgact tccggcttca
2101 gtttggacga accaacttcc tttgcatcaa gaattaacag attgatctct ttaggcctga
2161 acattgatga ggatgaagaa acagagactg ctccagaagc atccaccgca gctccggttg
2221 aagaggttcc agctgacacc gaaatggaag aggtagatta atggatccga attcgagctc
2281 cgtcgacaag cttgcggccg cactcgagtc tggtaaagaa accgctgctg cgaaatttga
2341 acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc
2401 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
2461 gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt
2521 aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc
2581 gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcaa
2641 ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca
2701 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
2761 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
2821 atagtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
2881 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
2941 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
3001 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
3061 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
3121 aactccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
3181 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
3241 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
3301 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
3361 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
3421 gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg
3481 gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt
3541 tattcggcgc aaagtgcgtc gggtgatgct gccaacttac tgatttagtg tatgatggtg
3601 tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctcctgtt cagctactga
3661 cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctagcgga gtgtatactg
3721 gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa
3781 aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca
3841 ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg
3901 gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa
3961 gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca
4021 gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct
4081 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
4141 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
4201 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
4261 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
4321 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
4381 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
4441 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
4501 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
4561 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
4621 tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
4681 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
4741 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
4801 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
4861 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
4921 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
4981 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
5041 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
5101 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
5161 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
5221 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
5281 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
5341 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
5401 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
5461 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
5521 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
5581 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
5641 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
5701 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
5761 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
5821 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
5881 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattag
//
