LOCUS       sequence-149101-        6592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2793 is a vector for multiple integrations at sites sharing 
            homology with Ty2Cons. 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 6592)
  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 6592)
  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: pCfB2793
FEATURES             Location/Qualifiers
     source          1..6592
                     /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(2166..2199)
                     /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(2669..2685)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2784..2801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2955..3543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3035..3054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3667..4458,4459..4527))
                     /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     4290..4309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4528..4619)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4661..4680
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4686..4705
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4912..4945
                     /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(5226..5391)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5312..5336
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5440..5846
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5868..6581
                     /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(5898..5926)
                     /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 tcaatataaa
     1981 atgatgataa taatatttat agaattgtgt agaattgcag attccctttt atggattcct
     2041 aaatcctgag gagaacttct agtatattct acatacctaa tattattgcc ttattaaaaa
     2101 tggaatccca acaattacat caaaatccac attctcagat ctgcggccgc actcagacct
     2161 gaagtgaagt tcctatactt tctagagaat aggaacttct atagtgagtc gaataagggc
     2221 gacacaaaat ttattctaaa tgcataataa atactgataa catcttatag tttgtattat
     2281 attttgtatt atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta
     2341 ttttcgagat ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa
     2401 atcataaata atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta
     2461 tcttatttaa agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca
     2521 agcaaagtga caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca
     2581 taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc
     2641 agaaccgccc agggggcccg agcttaagac tggccgtcgt tttacaacac agaaagagtt
     2701 tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc ttagtttgat gcctggcagt
     2761 tccctactct cgccttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2821 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2881 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2941 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3001 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3061 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3121 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3181 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3241 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3301 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3361 ttgaagtggt gggctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3421 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3481 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3541 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgcgcgta
     3601 actcacgtta agggattttg gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct
     3661 ctgcttttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3721 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3781 tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg atttatcagc
     3841 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3901 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3961 gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4021 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4081 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4141 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4201 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4261 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4321 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4381 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4441 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4501 ggcgacacgg aaatgttgaa tactcatatt cttccttttt caatattatt gaagcattta
     4561 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4621 aggggtcagt gttacaacca attaaccaat tctgaacatt atcgcgagcc catttatacc
     4681 tgaatatggc tcataacacc ccttgtttgc ctggcggcag tagcgcggtg gtcccacctg
     4741 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggactcccc
     4801 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     4861 gcctttcgcc cgggctaatt atggggtgtc gcccttattc gactctatag tgaagttcct
     4921 attctctaga aagtatagga acttctgaag tggggattta aagtcggtgt ccgcgctgag
     4981 ggtttaatgg cgcgccgcgg ccgcccgcgg tgttggaata aaaatcaact atcatctact
     5041 aactagtatt tacgttacta gtatattatc atatacggtg ttagaagatg acgcaaatga
     5101 tgagaaatag tcatctaaat tagtggaagc tgaaacgcaa ggattgataa tgtaatagga
     5161 tcaatgaata ttaacgctag cattaagtcc tcagcgagca cgctgcttca tggaatgcgt
     5221 gcgatgagcg acctcatgct atacctgaga aagcaacctg acctacagga aagagttact
     5281 caagaataag aattttcgtt ttaaaaccta agagtcactt taaaatttgt atacacttat
     5341 tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg cttatttaga
     5401 agtgtcaaca acgtatctac caacggaatg cgtgcgatgc acacaccata gcttcaaaat
     5461 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
     5521 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
     5581 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
     5641 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat
     5701 ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct
     5761 caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga
     5821 aagaaagcat agcaatctaa tctaagtttt aattacaaat ctgtcatatg agcaaaggag
     5881 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     5941 acaaattttc tgtcagagga gagggtgaag gtgatgctac aatcggaaaa ctcaccctta
     6001 aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactctga
     6061 cctatggtgt tcaatgcttt tcccgttatc cggatcacat gaaaaggcat gactttttca
     6121 agagtgccat gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga
     6181 aatacaagac gcgtgctgta gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt
     6241 taaagggtac tgattttaaa gaagatggaa acattctcgg acacaaactc gagtacaact
     6301 ttaactcaca caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact
     6361 tcacagttcg ccacaacgtt gaagatggtt ccgttcaact agcagaccat tatcaacaaa
     6421 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaaa
     6481 ctgtcctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttcat gagtacgtaa
     6541 atgctgctgg gattacacat ggcatggatg agctctacaa atgaatcgcg tg
//
