LOCUS       sequence-149084-        7444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2798 is a vector for multiple integrations at sites sharing 
            homology with Ty4Cons. The selective marker is Kl.LEU2. Additionally
            it contains a PTEF1-GFP as reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7444)
  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 7444)
  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: pCfB2798
FEATURES             Location/Qualifiers
     source          1..7444
                     /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    2593..2626
                     /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(2669..2688)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2670..2688)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2698..2714)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2707..2729)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(3007..3040)
                     /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(3510..3526)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3625..3642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3796..4384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3876..3895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4508..5299,5300..5368))
                     /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     5131..5150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5369..5460)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5502..5521
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5527..5546
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5753..5786
                     /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(6078..6243)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6164..6188
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        6292..6698
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6720..7433
                     /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(6750..6778)
                     /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 agccaagatt tccttgacag
      481 ccttggcgat agcatcgcca acctcagtgg tagagttaga accaccaagg tcaccggttc
      541 tgacacctgc atccaagaca tttctaacag cttcttcaag agccctacct tcttcaacca
      601 aatccaagga taacttcaac atcatagctg cagataagat ggtagcaatt gggttaacct
      661 tgtttgctgg taaatctggg gcagaaccat gacatggttc gtacaaaccg aatgccttgt
      721 tagtgtcagg tagggaagct agagatgcag aaggtaataa acccaaagaa cctggaataa
      781 cagaggcttc atcggagata atatcaccaa acatgttgtt ggtaataaca acaccgttta
      841 gcttagttgg tgatttaacc aaaatcatag cagcagagtc gatcaattgg tgctgaacag
      901 ttaattgtgg gaactcagtc ttgatggttt cttcaacagt ctttctccac aatctggaag
      961 aggcaagcac gttagccttg tcaagtgacc agattggtaa tggtgggttt tgttgcaatg
     1021 ccaagaaagc agccattctt gtaattcttt gaacttcagg aacactgtat ttctcagagt
     1081 cccaagcaac tccgtcacct tcatcttctt ttctttcacc aaagtagata ccaccaacca
     1141 attctctaac gacgacgaaa tcggtaccct ttgcatattc aggcttcaaa ggagaaagat
     1201 ctagtaaaga atcagaagca aagttacatg gtcttaagtt ggcgtataga cccaattcct
     1261 ttctgatctt caatagacct tgttctggtc taactgcgcc cgtaccccat tttggaccac
     1321 caacagcacc tagtaagaca gcatcggctt tcttagaggc ttctagagct tcatctggta
     1381 aaggagtgcc agtggcatcg atggcagcac ccccgatcaa gtgatgttgg aaattgaact
     1441 taatttctgg acggacttca gcaatggcat tcaagacctt aatagcttcg tcagtaactt
     1501 ctttaccgac gtgatcaccc ggtaggacaa cgatattctt agacatggta taatctgtgt
     1561 agtgtgggat actttttact tcttcaaata ggtatcaaca ataaaaataa atcgaagcaa
     1621 atgtaggaat gcgttaaagc agatgtactt acttagagta catagatata tttatataat
     1681 tcaatatata aaagtatatg aagcatctga tgttgaacct gtcatgactc taaactggaa
     1741 tgggtagtta tgcttctgaa ggttttccgt tagaaaatga gtgcagggat caggcccgga
     1801 accggtttta gcctgaaagg aaaagaaaaa aatgcggtga ttactagcac gtgactgcgc
     1861 tgaattggaa tccaccacaa gtacagcgaa tggcaagagg gaaggggaat atattgtgcc
     1921 tggcggtagt ctgctatatc aaatgttaaa cataactaaa tgcagtagtg aatagagtaa
     1981 tgtatttcta catatgtggg tagggccaat gggagcccga tgtctattgt acagatattt
     2041 tccttttata gttaatcaat tcaacttgga aatcattgag tcgatatcta cttggataac
     2101 tggagccgtg gcagcgtttg aaatttgacg agtctccttg ttttgttcgc gcattttcag
     2161 tttattcaaa cgggagatgg caccgaatcc actgacttct ctgttcaatt tgaatggtaa
     2221 taggaagaaa gtacatttcg agaaattttc gcccaacgtc gaaatctcat ggatgatatc
     2281 ctcgatacaa atgacacctt catcacctaa tttctcttca acgatattgt tgtcattcaa
     2341 gattatctct tttggttctt tatcttctgg cctttgccac attattctag atctcttttg
     2401 gattaaagaa cggatagatg caagtgaagg ttgtccgata acaatatatg gtgatacaag
     2461 tttcaacaat gggtacacgt ccttggtaag tttgatgaaa acaccagtgt tcaactcaac
     2521 aagtctcaaa agagataaca ctttgtaagc ctttgctggg atcttcacag cgagctctcg
     2581 agaaccctta atataacttc gtataatgta tgctatacga agttattagg tgatatcaga
     2641 tccactagtg gcctatgcac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     2701 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttaccccta caggactagt
     2761 gctgaggcat taattgatca ggtaggtaca tatatgagga atatgagtcg tcacatcaat
     2821 gtatagtaac taccggaatc actattatat tggtcatgat taatatgacc aatcggcgtg
     2881 tgttttatat acctctctta tttagtataa gaagatcagt actcacttct tcattaatac
     2941 taatttttaa cctctaatta tcaacaagat ctgcggccgc ggccgcaaat ttaaataaaa
     3001 tgaagtgaag ttcctatact ttctagagaa taggaacttc tatagtgagt cgaataaggg
     3061 cgacacaaaa tttattctaa atgcataata aatactgata acatcttata gtttgtatta
     3121 tattttgtat tatcgttgac atgtataatt ttgatatcaa aaactgattt tccctttatt
     3181 attttcgaga tttattttct taattctctt taacaaacta gaaatattgt atatacaaaa
     3241 aatcataaat aatagatgaa tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt
     3301 atcttattta aagtgcgttg cttttttctc atttataagg ttaaataatt ctcatatatc
     3361 aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc tctttcccta aactcccccc
     3421 ataaaaaaac ccgccgaagc gggtttttac gttatttgcg gattaacgat tactcgttat
     3481 cagaaccgcc cagggggccc gagcttaaga ctggccgtcg ttttacaaca cagaaagagt
     3541 ttgtagaaac gcaaaaaggc catccgtcag gggccttctg cttagtttga tgcctggcag
     3601 ttccctactc tcgccttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4201 cttgaagtgg tgggctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgcgcgt
     4441 aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc
     4501 tctgctttta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4561 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4621 ctggccccag cgctgcgatg ataccgcgag aaccacgctc accggctccg gatttatcag
     4681 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4801 tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4921 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5101 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5161 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5341 gggcgacacg gaaatgttga atactcatat tcttcctttt tcaatattat tgaagcattt
     5401 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5461 taggggtcag tgttacaacc aattaaccaa ttctgaacat tatcgcgagc ccatttatac
     5521 ctgaatatgg ctcataacac cccttgtttg cctggcggca gtagcgcggt ggtcccacct
     5581 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggactccc
     5641 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     5701 ggcctttcgc ccgggctaat tatggggtgt cgcccttatt cgactctata gtgaagttcc
     5761 tattctctag aaagtatagg aacttctgaa gtggggattt aaatgcggcc gcgctgaggg
     5821 tttaatggcg cgccgcggcc gcccgcggtg ttggaacgag agtaattaat agtgacatga
     5881 gttgctatgg taacaatcta atgcttacat cgtatattaa tgtacaactc gtatacgttt
     5941 aagtgtgatt gcgcctattg cagaaggaat gttaaacgag aagctcagac aatactgaag
     6001 ctgtgttaaa gacctattag ttgaacatgt tatgctagca ttaagtcctc agcgagctcg
     6061 catggaatgc gtgcgatgag cgacctcatg ctatacctga gaaagcaacc tgacctacag
     6121 gaaagagtta ctcaagaata agaattttcg ttttaaaacc taagagtcac tttaaaattt
     6181 gtatacactt atttttttta taacttattt aataataaaa atcataaatc ataagaaatt
     6241 cgcttattta gaagtgtcaa caacgtatct accaacggaa tgcgtgcgat gcacacacca
     6301 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
     6361 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
     6421 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
     6481 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     6541 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
     6601 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
     6661 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa atctgtcata
     6721 tgagcaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     6781 atgttaatgg gcacaaattt tctgtcagag gagagggtga aggtgatgct acaatcggaa
     6841 aactcaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
     6901 tcactactct gacctatggt gttcaatgct tttcccgtta tccggatcac atgaaaaggc
     6961 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact atatctttca
     7021 aagatgacgg gaaatacaag acgcgtgctg tagtcaagtt tgaaggtgat acccttgtta
     7081 atcgtatcga gttaaagggt actgatttta aagaagatgg aaacattctc ggacacaaac
     7141 tcgagtacaa ctttaactca cacaatgtat acatcacggc agacaaacaa aagaatggaa
     7201 tcaaagctaa cttcacagtt cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc
     7261 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
     7321 tgtcgacaca aactgtcctt tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc
     7381 atgagtacgt aaatgctgct gggattacac atggcatgga tgagctctac aaatgaatcg
     7441 cgtg
//
