LOCUS       sequence-192469-        3571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Cerulean3 fused to the 5' of cDNA of the 
            fission yeast tropomyosin, cdc8..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3571)
  AUTHORS   Brooker HR, Geeves MA, Mulvihill DP
  TITLE     Analysis of biophysical and functional consequences of 
            tropomyosin-fluorescent protein fusions.
  JOURNAL   FEBS Lett. 2016 Sep;590(18):3111-21. doi: 10.1002/1873-3468.12346. 
            Epub 2016 Aug 23.
  PUBMED    27501521
REFERENCE   2  (bases 1 to 3571)
  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"; doi: 
            "10.1002/1873-3468.12346"; journalName: "FEBS Lett"; date: 
            "2016-09"; volume: "590"; issue: "18"; pages: "3111-21"
COMMENT     Sequence Label: pJC20Cerulean3-Cdc8
FEATURES             Location/Qualifiers
     source          1..3571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(200..910,911..913,914..916))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(242..263)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     591..610
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     850..871
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(926..943)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             954..976
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1007..1026)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1008..1026)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1111..1128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1282..1870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1362..1381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2041..2832,2833..2901))
                     /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     2664..2683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2902..3006)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3074..3092
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3137..3184)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3180..3198
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 cttctaactc ttcgagttga gactcggcgg cttcgctttt cctcgaaaga gactcatatt
       61 cttgctcttt aagagaaagt tgaagttcaa cttctttcaa cttggcttca gctgcttcag
      121 cacgagcgac agcctcatca gtctcagcac gagcggcatt aattttctct ctaagcttat
      181 ccattcctcc tcctcctccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
      241 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg
      301 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
      361 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgcag ttgagcccga
      421 agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg ccgtggatgg
      481 cgttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
      541 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
      601 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
      661 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccccagc
      721 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
      781 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
      841 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
      901 cgcccttgct caccattcct cctccatgat gatgatgatg atgcatatgt atatctcctt
      961 cttaaagtta aacaaaatta tttctagagg gaaaccgttg tggtctccct atagtgagtc
     1021 gtattaattt cgcgggatcg agatcctgca ttaatgaatc ggccaacgcg cggggagagg
     1081 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1141 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1201 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1681 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2161 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2641 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     2701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3001 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     3061 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat tatccggata
     3121 tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     3181 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
     3241 ccggatcagt tagctagggc ccgggatccc tacaaatcct caagagcttg gtgaacttca
     3301 tctaattcag ccttgacctt agtatacttg tcagtcattt cttctagttt ctgttccata
     3361 tcatcacgct cacgctctag agattggacg cgacgttcaa aatgctcagc cttaacatca
     3421 gtttgacgca tcttttcggt ggtttcgcga agtagcttgt cattagtttc tagttcttcc
     3481 tctaaaagct caactttacg gctcaactgc tcggcttccg tcttttgaat atcttcattg
     3541 tcggctttga gacgtaactg cttagtttct t
//
