LOCUS       sequence-192448-        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 
            Ala-18-Thr..
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.A18T
FEATURES             Location/Qualifiers
     source          1..2809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(199..990,991..1059))
                     /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     822..841
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1060..1164)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1232..1250
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1295..1342)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1338..1356
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             1921..1943
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1974..1993)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1975..1993)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2078..2095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2249..2809,1..28))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2329..2348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
       61 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      121 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      181 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      241 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
      301 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
      361 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
      421 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
      481 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      541 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      601 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
      661 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
      721 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
      781 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
      841 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
      901 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
      961 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1021 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     1081 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1141 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     1201 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     1261 tcaagaatta tccggatata gttcctcctt tcagcaaaaa acccctcaag acccgtttag
     1321 aggccccaag gggttatgct agttattgct cagcggtggc agcagccaac tcagcttcct
     1381 ttcgggcttt gttagcagcc ggatcagtta gctagggccc gggatcccta caaatcctca
     1441 agagcttggt gaacttcatc taattcagcc ttgaccttag tatacttgtc agtcatttct
     1501 tctagtttct gttccatatc atcacgctca cgctctagag attggacgcg acgttcaaaa
     1561 tgctcagcct taacatcagt ttgacgcatc ttttcggtgg tttcgcgaag tagcttgtca
     1621 ttagtttcta gttcttcctc taaaagctca actttacggc tcaactgctc ggcttccgtc
     1681 ttttgaatat cttcattgtc ggctttgaga cgtaactgct tagtttcttc ttctaactct
     1741 tcgagttgag actcggcggc ttcgcttttc ctcgaaagag actcatattc ttgctcttta
     1801 agagaaagtt gaagttcaac ttctttcaac ttggcttcag ctgcttcagc acgagcgaca
     1861 gtctcatcag tctcagcacg agcggcatta attttctctc taagcttatc catatgtata
     1921 tctccttctt aaagttaaac aaaattattt ctagagggaa accgttgtgg tctccctata
     1981 gtgagtcgta ttaatttcgc gggatcgaga tcctgcatta atgaatcggc caacgcgcgg
     2041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagca
//
