LOCUS       sequence-148938-        6605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2792 is a vector for multiple integrations at sites sharing 
            homology with Ty3Cons. The selective marker is Kl.URA3. Additionally
            it contains a PTEF1-GFP as reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6605)
  AUTHORS   Maury J, Germann SM, Baallal Jacobsen SA, Jensen NB, Kildegaard KR, 
            Herrgard MJ, Schneider K, Koza A, Forster J, Nielsen J, Borodina I
  TITLE     EasyCloneMulti: A Set of Vectors for Simultaneous and Multiple 
            Genomic Integrations in Saccharomyces cerevisiae.
  JOURNAL   PLoS One. 2016 Mar 2;11(3):e0150394. doi: 
            10.1371/journal.pone.0150394. eCollection 2016.
  PUBMED    26934490
REFERENCE   2  (bases 1 to 6605)
  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: "PLoS One.";
            date: "2016-03-2"; volume: "11(3):e0150394. doi"; pages: " 
            10.1371/journal.pone.0150394. eCollection 2016"
COMMENT     Sequence Label: pCfB2792
FEATURES             Location/Qualifiers
     source          1..6605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      6..195
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     protein_bind    243..276
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1777..1810
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1853..1872)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1854..1872)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1882..1898)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1891..1913)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(2174..2207)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(2677..2693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2792..2809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2963..3551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3043..3062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3675..4466,4467..4535))
                     /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     4298..4317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4536..4627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4669..4688
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4694..4713
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4920..4953
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     terminator      complement(5239..5404)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5325..5349
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5453..5859
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5881..6594
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEKRDHMVLHE
                     YVNAAGITHGMDELYK"
     primer_bind     complement(5911..5939)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 cattcatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
       61 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
      121 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
      181 ttgggacgct cgaagatcgc gtcagctgaa gcttcgtacg ctgcaggtcg acaaccctta
      241 atataacttc gtataatgta tgctatacga agttattagg tctagagatc ccaatacaac
      301 agatcacgtg atctttttgt aagatgaagt tgaagtagtg ttgcaccgtg ccaatgcagg
      361 tggctattag attaaatatg tgatttgttc tattaagttt cctgtataat tacaaatgaa
      421 taacgaaatg agacaaagaa gagaaccaat ttttacaagc atggggagcg ctgattctct
      481 tttggtacgc ttcccatcca gcatttctgt atctttcacc ttcaacctta ggatctctac
      541 ccttggcgaa aagtcctctg ccaacaatga tgatatctga tccaccactt acaacttcgt
      601 cgacggttct gtactgctga cccaatgcat cgcctttgtc gtctaaacct acacctgggg
      661 tcatgattag ccaatcaaac ccttcttctc ttcctcccat atcgttctga gcaatgaacc
      721 caataacgaa atctttatca ctctttgcaa tatcaacggt acccttagta tattcaccgt
      781 gtgctagaga acccttggaa gataattcag caagcatcaa taatcccctt ggttctttgg
      841 tgacctcttg cgcaccttgt ttcaagccag caacaatacc agcaccagta accccgtggg
      901 cgttggtgat atcagaccat tctgcgatac ggtaaacgcc cgatgtatat tgtaatttga
      961 ctgtgttacc gatatcggcg aattttctgt cctcaaatat caagaacttg tatttctctg
     1021 ccaatgcttt caatggaacg acagtaccct cataactgaa atcatccaag atatcaacgt
     1081 gtgttttcaa aaggcaaatg tatggaccca acgtttcaac aagtttcaat agctcatcag
     1141 tcgaacgaac gtcaagagaa gcacacaaat tggttttctt ttcatccatt aaacgtaaaa
     1201 gtttcgatgc aaccggactt gcatgagtct cagctctact ggtatatgat tttgtggaca
     1261 tggtgcaact aattgacggg agtgtattga cgctggcgta ctggctttca caaaatggcc
     1321 caatcacaac cacatcttag atagttgaaa tgactttaga taacatcaat tgagatgagc
     1381 ttaatcatgt caaagctaaa agtgtcacca tgaacgacaa ttcttaagca aatcacgtga
     1441 tatagatcca cgaataacca ccatttgatg ctcgaggcaa gtaatgtgtg taaaaaaatg
     1501 cgttaccacc atccaatgca gaccgatctt ctacccagaa tcacatatat ttatgtaccg
     1561 agtacctttt ttctatcttc caattgcttc tcccatatga ttgtctccgt aagctcgaaa
     1621 tttctaagtt ggattttaat cttcacgcag gatgacagtt cgatgagctt ctgaggagtg
     1681 tttagaacat aatcagttta tccatggtct atctcttctt gtcgcttttt ctcctcgata
     1741 gaacctaaat aaaacgagct ctcgagaacc cttaatataa cttcgtataa tgtatgctat
     1801 acgaagttat taggtgatat cagatccact agtggcctat gcacccaatt cgccctatag
     1861 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1921 tggcgttacc cctgcaggac agacgtcatt agtgctgagg cattaattga tcagattccg
     1981 cgcttccacc acttagtatg attcatattt tatataatat ataagataag taacattccg
     2041 tgaattaatc tgataaactg ttttgacaac tggttacttc cctaagactg tttatattag
     2101 gattgtcaag acactccggt attactcgag cccgtaatac aacaagatct gcggccgcac
     2161 tcagacctga agtgaagttc ctatactttc tagagaatag gaacttctat agtgagtcga
     2221 ataagggcga cacaaaattt attctaaatg cataataaat actgataaca tcttatagtt
     2281 tgtattatat tttgtattat cgttgacatg tataattttg atatcaaaaa ctgattttcc
     2341 ctttattatt ttcgagattt attttcttaa ttctctttaa caaactagaa atattgtata
     2401 tacaaaaaat cataaataat agatgaatag tttaattata ggtgttcatc aatcgaaaaa
     2461 gcaacgtatc ttatttaaag tgcgttgctt ttttctcatt tataaggtta aataattctc
     2521 atatatcaag caaagtgaca ggcgccctta aatattctga caaatgctct ttccctaaac
     2581 tccccccata aaaaaacccg ccgaagcggg tttttacgtt atttgcggat taacgattac
     2641 tcgttatcag aaccgcccag ggggcccgag cttaagactg gccgtcgttt tacaacacag
     2701 aaagagtttg tagaaacgca aaaaggccat ccgtcagggg ccttctgctt agtttgatgc
     2761 ctggcagttc cctactctcg ccttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2821 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2881 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2941 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3001 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3061 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3361 cagagttctt gaagtggtgg gctaactacg gctacactag aagaacagta tttggtatct
     3421 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgacg
     3601 cgcgcgtaac tcacgttaag ggattttggt catgagcttg cgccgtcccg tcaagtcagc
     3661 gtaatgctct gcttttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3781 ttaccatctg gccccagcgc tgcgatgata ccgcgagaac cacgctcacc ggctccggat
     3841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3961 aatagtttgc gcaacgttgt tgccatcgct acaggcatcg tggtgtcacg ctcgtcgttt
     4021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4501 ggaataaggg cgacacggaa atgttgaata ctcatattct tcctttttca atattattga
     4561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4621 aaacaaatag gggtcagtgt tacaaccaat taaccaattc tgaacattat cgcgagccca
     4681 tttatacctg aatatggctc ataacacccc ttgtttgcct ggcggcagta gcgcggtggt
     4741 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     4801 gactccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     4861 aagactgggc ctttcgcccg ggctaattat ggggtgtcgc ccttattcga ctctatagtg
     4921 aagttcctat tctctagaaa gtataggaac ttctgaagtg gggatttaaa gtcggtgtcc
     4981 gcgctgaggg tttaatggcg cgccgcggcc gcccgcggtg ttgtatctca aaatgagata
     5041 tgtcagtatg acaatacgtc atcctgaacg ttcataaaac acatatgaaa caaccttata
     5101 acaaaacgaa caacatgaga caaaacccgt ccttccctag ctgaactacc caaaagtata
     5161 aatgcctgaa caattagttt agatccgagc tagcattaag tcctcagcga gcacgctgct
     5221 tcatggaatg cgtgcgatga gcgacctcat gctatacctg agaaagcaac ctgacctaca
     5281 ggaaagagtt actcaagaat aagaattttc gttttaaaac ctaagagtca ctttaaaatt
     5341 tgtatacact tatttttttt ataacttatt taataataaa aatcataaat cataagaaat
     5401 tcgcttattt agaagtgtca acaacgtatc taccaacgga atgcgtgcga tgcacacacc
     5461 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
     5521 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
     5581 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
     5641 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
     5701 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
     5761 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
     5821 cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca aatctgtcat
     5881 atgagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
     5941 gatgttaatg ggcacaaatt ttctgtcaga ggagagggtg aaggtgatgc tacaatcgga
     6001 aaactcaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
     6061 gtcactactc tgacctatgg tgttcaatgc ttttcccgtt atccggatca catgaaaagg
     6121 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
     6181 aaagatgacg ggaaatacaa gacgcgtgct gtagtcaagt ttgaaggtga tacccttgtt
     6241 aatcgtatcg agttaaaggg tactgatttt aaagaagatg gaaacattct cggacacaaa
     6301 ctcgagtaca actttaactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
     6361 atcaaagcta acttcacagt tcgccacaac gttgaagatg gttccgttca actagcagac
     6421 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     6481 ctgtcgacac aaactgtcct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
     6541 catgagtacg taaatgctgc tgggattaca catggcatgg atgagctcta caaatgaatc
     6601 gcgtg
//
