LOCUS       sequence-106686-        4621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recipient replicative vector to construct FLAG tagged fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4621)
  AUTHORS   Yanez-Carrillo P, Orta-Zavalza E, Gutierrez-Escobedo G, 
            Patron-Soberano A, De Las Penas A, Castano I
  TITLE     Expression vectors for C-terminal fusions with fluorescent proteins 
            and epitope tags in Candida glabrata.
  JOURNAL   Fungal Genet Biol. 2015 May 15. pii: S1087-1845(15)00087-0. doi: 
            10.1016/j.fgb.2015.04.020.
  PUBMED    25986172
REFERENCE   2  (bases 1 to 4621)
  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: "Fungal 
            Genet Biol. 2015 May 15. pii: S1087-1845(15)00087-0. doi: 
            10.1016/j.fgb.2015.04.020."
COMMENT     Sequence Label: pYC64
FEATURES             Location/Qualifiers
     source          1..4621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        341..747
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             761..1333
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARELGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1340..1587
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1356..1374)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     1608..1624
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1609..1625
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1672..1695
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1702..1725
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1732..1755
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1824..1844)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1824..1842)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1863..1879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1863..1879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1876..1898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1887..1903
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1911..1917,1918..1935,1936..1941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1956..1977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2094..2111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2265..2853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2345..2364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3024..3815,3816..3884))
                     /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     3647..3666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3885..3956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4594..4612
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcacctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca taccataaag cacagctcca tagcttcaaa
      361 atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca
      421 cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt
      481 taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc
      541 gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt ttttttttga
      601 tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc
      661 tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag
      721 aaagaaagca tagcaatcta atctaagttt tggatctaaa atgggcacca ctcttgacga
      781 cacggcttac cgataccgca ccagtgtccc gggggacgcc gaggccatcg aggcactgga
      841 tgggtccttc accaccgaca ccgtcttccg cgtcaccgcc accggggacg gcttcaccct
      901 gcgggaggtg ccggtggacc cgcccctgac caaggtgttc cccgacgacg aatcggacga
      961 cgaatcggac gacggggagg acggcgaccc tgactcccgg acgttcgtcg cgtacgggga
     1021 cgacggcgac ctggcgggct tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac
     1081 cgtcgaggac atcgaggtcg ccccggagca ccgggggcac ggggtcgggc gcgcgttgat
     1141 ggggctcgcg acggagttcg cccgcgagct gggcgccggg cacctctggc tggaggtcac
     1201 caacgtcaac gcaccggcga tccacgcgta ccggcggatg gggttcaccc tctgcggcct
     1261 ggacaccgcc ctgtacgacg gcaccgcctc ggacggcgag caggcgctct acatgagcat
     1321 gccctgcccc taagtcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     1381 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1441 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1501 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     1561 ttgggacgct cgaaggcttt aatttgcggc cggtaccggg ccccccctcg aggtcgacgg
     1621 tatcgataag cttgatatcg aattcctgca gcccggggga tcctaggagc agactacaaa
     1681 gatgacgatg acaaaggagc cgattataag gacgatgacg ataaaggggc tgactacaaa
     1741 gatgacgatg acaagtgata agatccgaag ttcctatact ttctagagcg gccgccaccg
     1801 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     1861 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2701 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtggg
     3961 gatctctaat gatgcaattt ttgttttgta cactaagatt ctatttcgga actcttaccc
     4021 atgtcattat ttactttatt ttcattattg taatttttta agcttttatt atgaattatt
     4081 taaatacaat tctttttgat catgtgacag acatcactga aagatgtgct ctgcgttatt
     4141 agtcagattg aaatatacat tttcatgtag atatgatgtt cttactttaa atgttgttcc
     4201 atttgctagc ttaaatagac gatattataa atttaagtat tcactaaaaa tctgacccat
     4261 taatgttaga attagtatta ttttttgtga tattacattt ttatattttg gatttcaacc
     4321 aatgctctgc gatatgattc aacagaataa tatgtacaag ctactttaat tcatatatac
     4381 taatagattc acagatccag tacgactgca agaatttacg ttttgtttga aatggtagta
     4441 gctacactaa attaacaatg atcgattaat ccacatccac agtgtgtaat catcctaatg
     4501 aaaacaagaa taactacaaa aatgaatata aactttttgc atgttagtct aacctgctaa
     4561 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4621 c
//
