LOCUS       sequence-82820-e        4128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  chromosomal tagging with Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4128)
  AUTHORS   Onishi M, Ko N, Nishihama R, Pringle JR
  TITLE     Distinct roles of Rho1, Cdc42, and Cyk3 in septum formation and 
            abscission during yeast cytokinesis.
  JOURNAL   J Cell Biol. 2013 Jul 22;202(2):311-29. doi: 10.1083/jcb.201302001.
  PUBMED    23878277
REFERENCE   2  (bases 1 to 4128)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2013-07-22- 22"; volume: "202"; 
            issue: "2"; pages: "311-29"
COMMENT     Sequence Label: pFA6a-VN155(I153L)-TRP1
FEATURES             Location/Qualifiers
     source          1..4128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      70..79
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(76..78,79..81,82..540)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYLTA"
     primer_bind     complement(121..142)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(382..401)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     terminator      564..751
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(619..643)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        787..934
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             935..1609
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     RBS             1683..1691
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1763..1782)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1764..1782)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1869..1886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2040..2628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2120..2139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2799..3590,3591..3659))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3422..3441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3660..3764)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3832..3850
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3888..3910)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4010..4029
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4110..4128
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4110..4127
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 agatccatcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      121 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      181 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
      241 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
      301 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      361 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      421 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      481 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tctcaccgcc
      541 tagggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
      601 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
      661 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg
      721 aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat
      781 agatctgtac aatcttgatc cggagctttt ctttttttgc cgattaagaa ttaattcggt
      841 cgaaaaaaga aaaggagagg gccaagaggg agggcattgg tgactattga gcacgtgagt
      901 atacgtgatt aagcacacaa aggcagcttg gagtatgtct gttattaatt tcacaggtag
      961 ttctggtcca ttggtgaaag tttgcggctt gcagagcaca gaggccgcag aatgtgctct
     1021 agattccgat gctgacttgc tgggtattat atgtgtgccc aatagaaaga gaacaattga
     1081 cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca tataaaaata gttcaggcac
     1141 tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag gaggatgttt tggctctggt
     1201 caatgattac ggcattgata tcgtccaact gcatggagat gagtcgtggc aagaatacca
     1261 agagttcctc ggtttgccag ttattaaaag actcgtattt ccaaaagact gcaacatact
     1321 actcagtgca gcttcacaga aacctcattc gtttattccc ttgtttgatt cagaagcagg
     1381 tgggacaggt gaacttttgg attggaactc gatttctgac tgggttggaa ggcaagagag
     1441 ccccgaaagc ttacatttta tgttagctgg tggactgacg ccagaaaatg ttggtgatgc
     1501 gcttagatta aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt
     1561 aaaagactct aacaaaatag caaatttcgt caaaaatgct aagaaatagg ttattactga
     1621 gtagtattta tttaagtatt gtttgtgcac ttgcctgcgg tgtgaaatac cgcacagatg
     1681 cgtaaggagg tttaaacgag ctcgaattca tcgatgatat cagatccact agtggcctat
     1741 gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat aagccaggtt
     1801 aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1861 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1921 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1981 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2041 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2101 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2161 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2221 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2281 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2341 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2401 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2461 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2521 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2581 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2641 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2701 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2761 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2821 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2881 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2941 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3001 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3061 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3121 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3181 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3241 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3301 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3361 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3421 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3481 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3541 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3601 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3661 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3721 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3781 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     3841 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3901 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3961 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4021 cagattgtac tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa
     4081 tacataacct tatgtatcat acacatacga tttaggtgac actataga
//
