LOCUS       sequence-192456-        2809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector containing cDNA encoding for the 
            temperature sensitive fission yeast tropomyosin mutant, Cdc8.110..
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: pJC20cdc8.110
FEATURES             Location/Qualifiers
     source          1..2809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(126..714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(206..225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(885..1676,1677..1745))
                     /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     1508..1527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1746..1850)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1918..1936
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1981..2028)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2024..2042
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             2607..2629
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2660..2679)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2661..2679)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2764..2781)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
       61 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      541 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1501 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1861 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     1921 aggcgtatca cgaggccctt tcgtcttcaa gaattatccg gatatagttc ctcctttcag
     1981 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc
     2041 ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat cagttagcta
     2101 gggcccggga tccctacaaa tcctcaagag cttggtgaac ttcatctaat tcagccttga
     2161 ccttagtata cttgtcagtc atttcttcta gtttctgttc catatcatca cgctcacgct
     2221 ctagagattg gacgcgacgt tcaaaatgct cagccttaac atcagtttga cgcatctttt
     2281 cggtggtttc gcgaagtagc ttgtcattag tttctagttc ttcctctaaa agctcaactt
     2341 tacggctcaa ctgctcggct tccgtctttt gaatatcttc attgtcggct ttgagacgta
     2401 actgcttagt ttcttcttct aactcttcga gttgagactc ggcggcttcg cttttcctcg
     2461 aaagagactc atattcttgc tctttaagag aaagttgaag ttcaactttt ttcaacttgg
     2521 cttcagctgc ttcagcacga gcgacagtct catcagtctc agcacgagcg gcattaattt
     2581 tctctctaag cttatccata tgtatatctc cttcttaaag ttaaacaaaa ttatttctag
     2641 agggaaaccg ttgtggtctc cctatagtga gtcgtattaa tttcgcggga tcgagatcct
     2701 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2761 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcg
//
