LOCUS       sequence-391042-        5937 bp    DNA     circular SYN 14-APR-2025
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 Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.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"
COMMENT     Sequence Label: pACYC-T7
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
//
