LOCUS       sequence-228928-        8133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8133)
  AUTHORS   Shorter J, Lindquist S
  TITLE     Hsp104 catalyzes formation and elimination of self-replicating Sup35
            prion conformers.
  JOURNAL   Science 2004 Jun 18;304(5678):1793-7. Epub 2004 May 20.
  PUBMED    15155912
REFERENCE   2  (bases 1 to 8133)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science"; 
            date: "2004-06-18"; volume: "304(5678)"; pages: "1793-7. Epub 2004 
            May 20"
COMMENT     Sequence Label: 6107 pETH6KT218
FEATURES             Location/Qualifiers
     source          1..8133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2998..3015
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3068..3086)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3082..3129
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3166..3621
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3253..3272)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3463..3484
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3713..4528)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3713..4528)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4436..4455
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4520..4539
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4650..5238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5139..5158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5392..5409
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5424..5563
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5489..5508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5649..5671
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5665..5856)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6665..7747)
                     /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     7709..7728
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7748..7825)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     8031..8050
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg aacgaccaaa cgcaatttac agaaagggct
      181 ctaacgattt tgacgttggc tcaaaaattg gcttcggatc atcaacatcc acaattacaa
      241 cctatacata ttctagctgc cttcattgaa acgccagaag atggatcagt cccttaccta
      301 cagaatctaa ttgagaaggg ccgttacgac tatgatcttt tcaagaaagt ggttaataga
      361 aatctagtaa gaattcctca acagcaacct gcacctgcgg agataactcc aagttatgct
      421 ttggggaaag tccttcaaga cgctgctaag attcaaaaac aacagaagga ctcatttata
      481 gcgcaagacc atatattgtt tgctctattc aatgattcgt ctattcagca aatatttaag
      541 gaagctcaag tagatattga ggccatcaag caacaagctc ttgaacttcg tggtaacact
      601 agaattgact ctcgtggcgc tgatacgaac acacctttgg aatatttatc aaagtacgcc
      661 attgatatga ctgagcaggc tcgtcaaggt aaacttgacc ctgtcatcgg ccgtgaagaa
      721 gaaataagaa gcactattag agttttagca agaagaatta agtccaaccc atgtttaatt
      781 ggtgagccag gtatcggtac caccgctatt attgaaggtg ttgctcaaag aatcattgac
      841 gatgacgttc ccactatctt acaaggcgct aaattgttca gtctagattt ggccgcatta
      901 accgcaggtg ctaaatacaa aggtgatttc gaagaaagat tcaaaggtgt tttgaaggaa
      961 atcgaagaat caaagactct aattgtgtta ttcattgatg aaattcacat gttaatgggt
     1021 aatggtaagg acgacgctgc taacatcttg aagccagctt tgtccagagg ccaattgaag
     1081 gtcatcggtg ccaccaccaa taacgaatat agatctattg tggaaaagga tggtgccttt
     1141 gaaagaagat tccagaaaat tgaagtcgct gaaccaagtg tgagacaaac agtggccata
     1201 ttgagaggtc tgcaaccaaa gtatgaaata catcatggtg taaggattct ggatagcgcc
     1261 ttagtcactg ctgctcaatt agccaagcgt tacttgccat atagaagatt gccagattct
     1321 gctttggatt tagttgatat ttcttgtgct ggtgtcgccg tcgcaagaga ttctaagcca
     1381 gaagaattgg attccaagga acgtcaattg caattgattc aagtagagat aaaagctcta
     1441 gagagagatg aagatgccga ctccaccact aaagatagat taaagttagc taggcagaag
     1501 gaagcttcat tgcaagaaga attggaacct ctaagacaac gttacaatga agaaaagcat
     1561 ggccatgaag aattgacaca agctaaaaag aaattggatg aactggaaaa caaggccctt
     1621 gatgctgaac gtagatatga tactgctacc gccgctgatt taaggtactt cgccatccca
     1681 gatatcaaaa agcaaatcga aaagcttgaa gatcaggttg ctgaggaaga gagacgtgct
     1741 ggtgccaact ccatgatcca aaatgtggtc gattcagaca ccatttctga aacagctgca
     1801 agattgactg gtatccctgt taagaagttg tcagaatctg aaaatgaaaa attgattcat
     1861 atggaacgtg acttatcatc tgaagtcgtg ggccaaatgg atgccattaa agctgtttcc
     1921 aatgccgtta gattgtctag atcaggttta gctaatccaa ggcaaccagc atccttctta
     1981 tttttaggtt tgtccggttc cggtaaaact gaattggcta aaaaagttgc tggatttttg
     2041 tttaatgatg aggacatgat gatcagggtc gattgttctg aattaagcga gaagtatgcg
     2101 gtctctaagt tgttgggtac cacggcaggt tatgtcgggt acgatgaagg tggcttttta
     2161 actaaccaac tgcaatacaa accatactcc gttttgttat tcgatgaagt agaaaaggca
     2221 catcctgatg ttttgactgt catgctacaa atgttggatg acggtagaat tacttctggt
     2281 caaggtaaga cgatcgactg ttccaattgt attgtcatca tgacttccaa tctaggtgct
     2341 gaatttatca attctcaaca aggatcaaag atccaagaat ctaccaagaa tttggtcatg
     2401 ggtgctgtta ggcaacattt cagaccagaa tttttgaaca gaatttctag tatagtcatt
     2461 ttcaacaagc tatctagaaa agctattcat aagatcgtgg atattcgttt gaaggaaatt
     2521 gaagagagat tcgagcaaaa tgataaacat tacaagttga atttaactca agaggccaag
     2581 gacttcttgg ccaaatatgg ttattccgat gatatgggtg cacgtccact gaacaggtta
     2641 attcaaaacg aaattttgaa caaactggca ctaaggatct taaagaatga aatcaaggat
     2701 aaggaaactg tcaatgtcgt cttgaagaag ggtaaatctc gtgatgaaaa tgttcctgag
     2761 gaagctgaag aatgtctgga agttctacca aatcacgaag ctactatagg ggctgacacg
     2821 ttaggtgatg acgataatga ggacagtatg gaaattgatg atgacctaga ttaatttaat
     2881 atagtgtgat ttttaaaaac tttcgaacaa gaatcagtaa tataatatat ataattaata
     2941 aaactaatgg aatttgttta attgaacttg acacccgagc aaaagagctc cgtcgagcac
     3001 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     3061 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     3121 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
     3181 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3241 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3301 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3361 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3421 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3481 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3541 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3601 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3661 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     3721 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     3781 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     3841 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     3901 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     3961 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     4021 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     4081 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     4141 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     4201 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     4261 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     4321 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     4381 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     4441 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     4501 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     4561 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4621 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4681 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4741 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4801 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4861 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4921 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4981 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5041 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5101 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5161 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5221 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5281 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5341 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5401 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5461 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5521 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     5581 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5641 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5701 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     5761 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     5821 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     5881 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     5941 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     6001 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     6061 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     6121 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     6181 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     6241 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     6301 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     6361 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     6421 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     6481 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     6541 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     6601 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     6661 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6721 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     6781 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     6841 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     6901 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     6961 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     7021 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     7081 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     7141 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     7201 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     7261 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     7321 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     7381 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     7441 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     7501 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     7561 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     7621 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     7681 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     7741 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     7801 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     7861 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     7921 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     7981 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     8041 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     8101 gcgtagagga tcgagatctc gatcccgcga aat
//
