LOCUS       sequence-192453-        2809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector containing cDNA encoding for the 
            temperature sensitive fission yeast tropomyosin mutant, Cdc8.27..
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: pJC20cdc8.27
FEATURES             Location/Qualifiers
     source          1..2809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     350..369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     603..620
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     705..724
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        705..723
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             755..777
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1342..1360)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1356..1403
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1448..1466)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1534..1638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1639..1707,1708..2499)
                     /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(1857..1876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2670..2809,1..449)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
       61 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
      121 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
      181 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      241 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      301 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      361 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      421 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      481 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      541 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      601 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
      661 cgcgttggcc gattcattaa tgcaggatct cgatcccgcg aaattaatac gactcactat
      721 agggagacca caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat
      781 acatatggat aagcttagag agaaaattaa tgccgctcgt gctgagactg atgaggctgt
      841 cgctcgtgct gaagcagctg aagccaagtt gaaagaagtt gaacttcaac tttctcttaa
      901 agagcaagaa tatgagtctc tttcgaggaa aagcgaagcc gccgagtctc aactcgaaga
      961 gttagaagaa gaaactaagc agttacgtct caaagccgac aatgaagata ttcaaaagac
     1021 ggaagccgag cagttgagcc gtaaagttga gcttttagag gaagaactag aaactaatga
     1081 caagctactt cgcgaaacca ccgaaaagat gcgtcaaact gatgttaagg ctgagcattt
     1141 tgaacgtcgc gtccaatctc tagagcgtaa gcgtgatgat atggaacaga aactagaaga
     1201 aatgactgac aagtatacta aggtcaaggc tgaattagat gaagttcacc aagctcttga
     1261 ggatttgtag ggatcccggg ccctagctaa ctgatccggc tgctaacaaa gcccgaaagg
     1321 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1381 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggataa ttcttgaaga
     1441 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1501 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1561 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1621 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1681 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1741 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1801 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1861 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1921 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1981 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2041 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2101 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2161 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2221 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2281 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2341 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2401 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2461 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2521 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2581 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2641 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2761 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaa
//
