LOCUS       sequence-181983-        2807 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 2807)
  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 2807)
  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..2807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..94
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        75..93
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             125..147
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(711..729)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      725..772
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(816..834)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        902..1006
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1007..1075,1076..1867)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1225..1244)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2038..2626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2780..2797
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct
       61 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaaa
      121 taattttgtt taactttaag aaggagatgt acatatggat aagcttagag agaaaattaa
      181 tgccgctcgt gctgagactg atgagactgt cgctcgtgct gaagcagctg aagccaagtt
      241 gaaaaaagtt gaacttcaac tttctcttaa agagcaagaa tatgagtctc tttcgaggaa
      301 aagcgaagcc gccgagtctc aactcgaaga gttagaagaa gaaactaagc agttacgtct
      361 caaagccgac aatgaagata ttcaaaagac ggaagccgag cagttgagcc gtaaagttga
      421 gcttttagag gaagaactag aaactaatga caagctactt cgcgaaacca ccgaaaagat
      481 gcgtcaaact gatgttaagg ctgagcattt tgaacgtcgc gtccaatctc tagagcgtga
      541 gcgtgatgat atggaacaga aactagaaga aatgactgac aagtatacta aggtcaaggc
      601 tgaattagat gaagttcacc aagctcttga ggatttgtag ggatcccggg ccctagctaa
      661 ctgatccggc tgctaacaaa gcccaaagga agctgagttg gctgctgcca ccgctgagca
      721 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
      781 aggaactata gataattctt cttgaagacg aaagggcctc gtgatacgcc tatttttata
      841 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
      901 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      961 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1021 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1081 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1141 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1201 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1261 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1321 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1381 aagcatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1441 gctaaccgct ttttttcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1501 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1561 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1621 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1681 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1741 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cgggcagtca
     1801 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1861 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1921 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1981 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2041 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2101 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2161 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2221 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2281 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2341 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2401 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     2461 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2521 tccagggggg aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2581 gcgtcgattt ttgtgatgct cgtcaggggg gccgagccta tggaaaaacg ccagcaacgc
     2641 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2701 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2761 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
