LOCUS       sequence-203674-        4882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast gene targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4882)
  AUTHORS   Wu JQ, Kuhn JR, Kovar DR, Pollard TD
  TITLE     Spatial and temporal pathway for assembly and constriction of the 
            contractile ring in fission yeast cytokinesis.
  JOURNAL   Dev Cell. 2003 Nov;5(5):723-34.
  PUBMED    14602073
REFERENCE   2  (bases 1 to 4882)
  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: "Dev Cell.";
            date: "2003-11"; volume: "5(5)"; pages: "723-34"
COMMENT     Sequence Label: pFA6a- mYFP-kanMX6
FEATURES             Location/Qualifiers
     source          1..4882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..40
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     22..39
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             101..814
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(128..156)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     751..771
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      835..1022
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(890..914)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1097..2453
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1097..1440
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1441..2250
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1508..1527)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2256..2453
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2557..2576)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2558..2576)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2663..2680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2834..3422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2914..2933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3593..4384,4385..4453))
                     /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     4216..4235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4454..4558)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4626..4644
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4682..4704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4804..4823
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
       61 ttcgtacgct gcaggtcgac ggatccccgg gttaattaac agtaaaggag aagaactttt
      121 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
      181 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg
      241 cactactgga aaactacctg ttccatggcc aacacttgtc actactttcg gttatggtct
      301 aaaatgcttt gctagatacc cagatcatat gaaacggcat gactttttca agagtgccat
      361 gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac
      421 acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
      481 tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca
      541 caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact tcaaaattag
      601 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
      661 tggcgatggc cctgtccttt taccagacaa ccattacctg tcctatcaat ctaagctttc
      721 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
      781 gattacacat ggcatggatg aactatacaa atagggcgcg ccacttctaa ataagcgaat
      841 ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa
      901 ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg
      961 taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc
     1021 ggcagatccg ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc
     1081 gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg
     1141 cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata
     1201 atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc
     1261 gctgcagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc
     1321 cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat
     1381 acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc
     1441 atgggtaagg aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat
     1501 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
     1561 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     1621 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     1681 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     1741 ggcaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     1801 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     1861 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     1921 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     1981 cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2041 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2101 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2161 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2221 tttcatttga tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt
     2281 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     2341 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     2401 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     2461 ttcgatacta acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc
     2521 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt
     2581 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2641 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3241 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3361 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4201 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4561 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4621 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     4681 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4741 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4801 atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac
     4861 gcggctacaa ttaatacata ac
//
