LOCUS       sequence-494945-        5241 bp    DNA     circular SYN 09-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5241)
  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 5241)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 9, 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: 6380 H10-K620T
FEATURES             Location/Qualifiers
     source          1..5241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     267..286
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        267..285
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    286..310
                     /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             325..347
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             360..389
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             402..413
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3325..3343)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3339..3386
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3431..3449)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3517..3621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3622..3690,3691..4482)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3840..3859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4653..5241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5142..5161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      241 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
      301 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggcc
      361 atcatcatca tcatcatcat catcatcaca gcagcggcca tatcgaaggt cgtcatatga
      421 acgaccaaac gcaatttaca gaaagggctc taacgatttt gacgttggct caaaaattgg
      481 cttcggatca tcaacatcca caattacaac ctatacatat tctagctgcc ttcattgaaa
      541 cgccagaaga tggatcagtc ccttacctac agaatctaat tgagaagggc cgttacgact
      601 atgatctttt caagaaagtg gttaatagaa atctagtaag aattcctcaa cagcaacctg
      661 cacctgcgga gataactcca agttatgctt tggggaaagt ccttcaagac gctgctaaga
      721 ttcaaaaaca acagaaggac tcatttatag cgcaagacca tatattgttt gctctattca
      781 atgattcgtc tattcagcaa atatttaagg aagctcaagt agatattgag gccatcaagc
      841 aacaagctct tgaacttcgt ggtaacacta gaattgactc tcgtggcgct gatacgaaca
      901 cacctttgga atatttatca aagtacgcca ttgatatgac tgagcaggct cgtcaaggta
      961 aacttgaccc tgtcatcggc cgtgaagaag aaataagaag cactattaga gttttagcaa
     1021 gaagaattaa gtccaaccca tgtttaattg gtgagccagg tatcggtaag accgctatta
     1081 ttgaaggtgt tgctcaaaga atcattgacg atgacgttcc cactatctta caaggcgcta
     1141 aattgttcag tctagatttg gccgcattaa ccgcaggtgc taaatacaaa ggtgatttcg
     1201 aagaaagatt caaaggtgtt ttgaaggaaa tcgaagaatc aaagactcta attgtgttat
     1261 tcattgatga aattcacatg ttaatgggta atggtaagga cgacgctgct aacatcttga
     1321 agccagcttt gtccagaggc caattgaagg tcatcggtgc caccaccaat aacgaatata
     1381 gatctattgt ggaaaaggat ggtgcctttg aaagaagatt ccagaaaatt gaagtcgctg
     1441 aaccaagtgt gagacaaaca gtggccatat tgagaggtct gcaaccaaag tatgaaatac
     1501 atcatggtgt aaggattctg gatagcgcct tagtcactgc tgctcaatta gccaagcgtt
     1561 acttgccata tagaagattg ccagattctg ctttggattt agttgatatt tcttgtgctg
     1621 gtgtcgccgt cgcaagagat tctaagccag aagaattgga ttccaaggaa cgtcaattgc
     1681 aattgattca agtagagata aaagctctag agagagatga agatgccgac tccaccacta
     1741 aagatagatt aaagttagct aggcagaagg aagcttcatt gcaagaagaa ttggaacctc
     1801 taagacaacg ttacaatgaa gaaaagcatg gccatgaaga attgacacaa gctaaaaaga
     1861 aattggatga actggaaaac aaggcccttg atgctgaacg tagatatgat actgctaccg
     1921 ccgctgattt aaggtacttc gccatcccag atatcaaaaa gcaaatcgaa aagcttgaag
     1981 atcaggttgc tgaggaagag agacgtgctg gtgccaactc catgatccaa aatgtggtcg
     2041 attcagacac catttctgaa acagctgcaa gattgactgg tatccctgtt aagaagttgt
     2101 cagaatctga aaatgaaaaa ttgattcata tggaacgtga cttatcatct gaagtcgtgg
     2161 gccaaatgga tgccattaaa gctgtttcca atgccgttag attgtctaga tcaggtttag
     2221 ctaatccaag gcaaccagca tccttcttat ttttaggttt gtccggttcc ggtaccactg
     2281 aattggctaa aaaagttgct ggatttttgt ttaatgatga ggacatgatg atcagggtcg
     2341 attgttctga attaagcgag aagtatgcgg tctctaagtt gttgggtacc acggcaggtt
     2401 atgtcgggta cgatgaaggt ggctttttaa ctaaccaact gcaatacaaa ccatactccg
     2461 ttttgttatt cgatgaagta gaaaaggcac atcctgatgt tttgactgtc atgctacaaa
     2521 tgttggatga cggtagaatt acttctggtc aaggtaagac gatcgactgt tccaattgta
     2581 ttgtcatcat gacttccaat ctaggtgctg aatttatcaa ttctcaacaa ggatcaaaga
     2641 tccaagaatc taccaagaat ttggtcatgg gtgctgttag gcaacatttc agaccagaat
     2701 ttttgaacag aatttctagt atagtcattt tcaacaagct atctagaaaa gctattcata
     2761 agatcgtgga tattcgtttg aaggaaattg aagagagatt cgagcaaaat gataaacatt
     2821 acaagttgaa tttaactcaa gaggccaagg acttcttggc caaatatggt tattccgatg
     2881 atatgggtgc acgtccactg aacaggttaa ttcaaaacga aattttgaac aaactggcac
     2941 taaggatctt aaagaatgaa atcaaggata aggaaactgt caatgtcgtc ttgaagaagg
     3001 gtaaatctcg tgatgaaaat gttcctgagg aagctgaaga atgtctggaa gttctaccaa
     3061 atcacgaagc tactataggg gctgacacgt taggtgatga cgataatgag gacagtatgg
     3121 aaattgatga tgacctagat taatttaata tagtgtgatt tttaaaaact ttcgaacaag
     3181 aatcagtaat ataatatata taattaataa aactaatgga atttgtttaa ttgaacttga
     3241 cacccgagca aaagagctcg gggccctagc taactgatcc ggctgctaac aaagcccgaa
     3301 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     3361 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga taattcttga
     3421 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3481 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3541 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3601 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3661 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3721 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3781 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3841 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     3901 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3961 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4021 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4081 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4141 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4201 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4261 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4321 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4381 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4441 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4501 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4561 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4621 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4681 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4741 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4801 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4861 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4921 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4981 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5041 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5101 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5161 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5221 ggggggcgga gcctatggaa a
//
