LOCUS       sequence-148932-        6601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2794 is a vector for multiple integrations at sites sharing 
            homology with Ty1Cons2. 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 6601)
  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 6601)
  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: pCfB2794
FEATURES             Location/Qualifiers
     source          1..6601
                     /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(2172..2205)
                     /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(2675..2691)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2790..2807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2961..3549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3041..3060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3673..4464,4465..4533))
                     /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     4296..4315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4534..4625)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4667..4686
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4692..4711
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4918..4951
                     /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(5235..5400)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5321..5345
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5449..5855
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5877..6590
                     /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(5907..5935)
                     /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 tcataaaacg
     1981 gaatgatgaa taatatttat agaattgtgt agaattgcag attccctttt atggattcct
     2041 aaatcctcga ggagaacttc tagtatattc tgtataccta atattatagc ctttatcaac
     2101 aatggaatcc caacaattat ctcaaaattc acatatttct caagatctgc ggccgcactc
     2161 agacctgaag tgaagttcct atactttcta gagaatagga acttctatag tgagtcgaat
     2221 aagggcgaca caaaatttat tctaaatgca taataaatac tgataacatc ttatagtttg
     2281 tattatattt tgtattatcg ttgacatgta taattttgat atcaaaaact gattttccct
     2341 ttattatttt cgagatttat tttcttaatt ctctttaaca aactagaaat attgtatata
     2401 caaaaaatca taaataatag atgaatagtt taattatagg tgttcatcaa tcgaaaaagc
     2461 aacgtatctt atttaaagtg cgttgctttt ttctcattta taaggttaaa taattctcat
     2521 atatcaagca aagtgacagg cgcccttaaa tattctgaca aatgctcttt ccctaaactc
     2581 cccccataaa aaaacccgcc gaagcgggtt tttacgttat ttgcggatta acgattactc
     2641 gttatcagaa ccgcccaggg ggcccgagct taagactggc cgtcgtttta caacacagaa
     2701 agagtttgta gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag tttgatgcct
     2761 ggcagttccc tactctcgcc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3361 gagttcttga agtggtgggc taactacggc tacactagaa gaacagtatt tggtatctgc
     3421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg
     3601 cgcgtaactc acgttaaggg attttggtca tgagcttgcg ccgtcccgtc aagtcagcgt
     3661 aatgctctgc ttttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3721 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3781 accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg ctccggattt
     3841 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3901 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3961 tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4021 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4081 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4141 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4201 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4261 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4321 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4381 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4441 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4501 aataagggcg acacggaaat gttgaatact catattcttc ctttttcaat attattgaag
     4561 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4621 acaaataggg gtcagtgtta caaccaatta accaattctg aacattatcg cgagcccatt
     4681 tatacctgaa tatggctcat aacacccctt gtttgcctgg cggcagtagc gcggtggtcc
     4741 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtgggga
     4801 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     4861 gactgggcct ttcgcccggg ctaattatgg ggtgtcgccc ttattcgact ctatagtgaa
     4921 gttcctattc tctagaaagt ataggaactt ctgaagtggg gatttaaagt cggtgtccgc
     4981 gctgagggtt taatggcgcg ccgcggccgc ccgcggtgtt ggaataaaaa tcaactatca
     5041 tctactaact agtatttacg ttactagtat attatcatat acggtgttag aagatgacgc
     5101 aaatgatgag aaatagtcat ctaaattagt ggaagctgaa acgcaaggat tgataatgta
     5161 ataggatcaa tgaatattaa catagctagc attaagtcct cagcgagcac gctgcttcat
     5221 ggaatgcgtg cgatgagcga cctcatgcta tacctgagaa agcaacctga cctacaggaa
     5281 agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt aaaatttgta
     5341 tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata agaaattcgc
     5401 ttatttagaa gtgtcaacaa cgtatctacc aacggaatgc gtgcgatgca cacaccatag
     5461 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc
     5521 cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg
     5581 gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt
     5641 tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt
     5701 ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct
     5761 tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg
     5821 ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaatc tgtcatatga
     5881 gcaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
     5941 ttaatgggca caaattttct gtcagaggag agggtgaagg tgatgctaca atcggaaaac
     6001 tcacccttaa atttatttgc actactggaa aactacctgt tccatggcca acacttgtca
     6061 ctactctgac ctatggtgtt caatgctttt cccgttatcc ggatcacatg aaaaggcatg
     6121 actttttcaa gagtgccatg cccgaaggtt atgtacagga acgcactata tctttcaaag
     6181 atgacgggaa atacaagacg cgtgctgtag tcaagtttga aggtgatacc cttgttaatc
     6241 gtatcgagtt aaagggtact gattttaaag aagatggaaa cattctcgga cacaaactcg
     6301 agtacaactt taactcacac aatgtataca tcacggcaga caaacaaaag aatggaatca
     6361 aagctaactt cacagttcgc cacaacgttg aagatggttc cgttcaacta gcagaccatt
     6421 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     6481 cgacacaaac tgtcctttcg aaagatccca acgaaaagcg tgaccacatg gtccttcatg
     6541 agtacgtaaa tgctgctggg attacacatg gcatggatga gctctacaaa tgaatcgcgt
     6601 g
//
