LOCUS       sequence-192449-        2809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector containing cDNA encoding for the fission
            yeast tropomyosin, Cdc8 with the amino acid substitution 
            Glu-31-Lys..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2809)
  AUTHORS   Johnson CA, Brooker HR, Gyamfi I, O'Brien J, Ashley B, Brazier JE, 
            Dean A, Embling J, Grimsey E, Tomlinson AC, Wilson EG, Geeves MA, 
            Mulvihill DP
  TITLE     Temperature sensitive point mutations in fission yeast tropomyosin 
            have long range effects on protein stability that destabilise the 
            actin-tropomyosin copolymer in vitro and in vivo.
  JOURNAL   Biochem Biophys Res Commun. 2017 Oct 25. pii: S0006-291X(17)32087-9.
            doi: 10.1016/j.bbrc.2017.10.109.
  PUBMED    29080743
REFERENCE   2  (bases 1 to 2809)
  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: "Biochem 
            Biophys Res Commun. 2017 Oct 25. pii: S0006-291X(17)32087-9. doi: 
            10.1016/j.bbrc.2017.10.109."
COMMENT     Sequence Label: pJC20cdc8.E31K
FEATURES             Location/Qualifiers
     source          1..2809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     312..331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     565..582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     667..686
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        667..685
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             717..739
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1304..1322)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1318..1365
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1410..1428)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1496..1600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1601..1669,1670..2461)
                     /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(1819..1838)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2632..2809,1..411)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
       61 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      481 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      541 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      601 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcaggat ctcgatcccg
      661 cgaaattaat acgactcact atagggagac cacaacggtt tccctctaga aataattttg
      721 tttaacttta agaaggagat atacatatgg ataagcttag agagaaaatt aatgccgctc
      781 gtgctgagac tgatgaggct gtcgctcgtg ctgaagcagc tgaagccaag ttgaaaaaag
      841 ttgaacttca actttctctt aaagagcaag aatatgagtc tctttcgagg aaaagcgaag
      901 ccgccgagtc tcaactcgaa gagttagaag aagaaactaa gcagttacgt ctcaaagccg
      961 acaatgaaga tattcaaaag acggaagccg agcagttgag ccgtaaagtt gagcttttag
     1021 aggaagaact agaaactaat gacaagctac ttcgcgaaac caccgaaaag atgcgtcaaa
     1081 ctgatgttaa ggctgagcat tttgaacgtc gcgtccaatc tctagagcgt gagcgtgatg
     1141 atatggaaca gaaactagaa gaaatgactg acaagtatac taaggtcaag gctgaattag
     1201 atgaagttca ccaagctctt gaggatttgt agggatcccg ggccctagct aactgatccg
     1261 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     1321 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     1381 atatccggat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     1441 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     1501 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1561 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1621 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1681 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1741 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1801 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     1861 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1921 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1981 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2041 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2101 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     2161 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2221 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2281 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2341 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2401 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2461 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2521 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2581 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2641 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2701 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2761 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccac
//
