LOCUS       sequence-494938-        4291 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 4291)
  AUTHORS   Glover JR, Lindquist S
  TITLE     Hsp104, Hsp70, and Hsp40: a novel chaperone system that rescues 
            previously aggregated proteins.
  JOURNAL   Cell 1998 Jul 10;94(1):73-82.
  PUBMED    9674429
REFERENCE   2  (bases 1 to 4291)
  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: "Cell"; 
            date: "1998-07-10"; volume: "94(1)"; pages: "73-82"
COMMENT     Sequence Label: 6379 H10-K218T
FEATURES             Location/Qualifiers
     source          1..4291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(584..603)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             595..1410
                     /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             595..1410
                     /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     complement(668..687)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2032..2109
                     /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             2110..3192
                     /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     complement(2129..2148)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3746..4291
                     /direction=RIGHT
                     /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     4192..4211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca tcttccagga
       61 aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg
      121 agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga cgcaccggtg
      181 cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag tgccaacata
      241 gtaagccagt atacactccg ctagcgctga ggtctgcctc gtgaagaagg tgttgctgac
      301 tcataccagg cctgaatcgc cccatcatcc agccagaaag tgagggagcc acggttgatg
      361 agagctttgt tgtaggtgga ccagttggtg attttgaact tttgctttgc cacggaacgg
      421 tctgcgttgt cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcgatttatt
      481 caacaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
      541 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
      601 catattcaac gggaaacgtc ttgctcgagg ccgcgattaa attccaacat ggatgctgat
      661 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
      721 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
      781 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
      841 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
      901 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
      961 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     1021 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     1081 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     1141 cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     1201 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     1261 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     1321 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     1381 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     1441 cagagcatta cgctgacttg acgggacggc ggctttgttg aataaatcga acttttgctg
     1501 agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa
     1561 agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca
     1621 ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca ccttcttcac
     1681 gaggcagacc tcagcgctca aagatgcagg ggtaaaagct aaccgcatct ttaccgacaa
     1741 ggcatccggc agttcaacag atcgggaagg gctggatttg ctgaggatga aggtggagga
     1801 aggtgatgtc attctggtga agaagctcga ccgtcttggc cgcgacaccg ccgacatgat
     1861 ccaactgata aaagagtttg atgctcaggg tgtagcggtt cggtttattg acgacgggat
     1921 cagtaccgac ggtgatatgg ggcaaatggt ggtcaccatc ctgtcggctg tggcacaggc
     1981 tgaacgccgg aggatccccg ggtaccgagc tcgaattcga gctcggtacc cgacaccatc
     2041 gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     2101 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     2161 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     2221 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     2281 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     2341 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     2401 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     2461 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     2521 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     2581 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     2641 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     2701 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     2761 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     2821 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     2881 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     2941 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     3001 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     3061 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     3121 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3181 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc
     3241 tttacacttt atgcttccga cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     3301 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3361 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3421 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3481 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3541 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3601 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3661 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3721 cagacccctt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     3781 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     3841 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     3901 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     3961 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     4021 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     4081 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     4141 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     4201 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     4261 atgcttgtca ggggggcgga gcctatggaa a
//
