LOCUS       sequence-148952-        6628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2791 is a vector for multiple integrations at sites sharing 
            homology with Ty4Cons. 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 6628)
  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 6628)
  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: pCfB2791
FEATURES             Location/Qualifiers
     source          1..6628
                     /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(2191..2224)
                     /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(2694..2710)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2809..2826)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2980..3568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3060..3079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3692..4483,4484..4552))
                     /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     4315..4334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4553..4644)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4686..4705
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4711..4730
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4937..4970
                     /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(5262..5427)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5348..5372
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5476..5882
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5904..6617
                     /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(5934..5962)
                     /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 cctacaggac tagtgctgag gcattaattg atcaggtagg tacatatatg
     1981 aggaatatga gtcgtcacat caatgtatag taactaccgg aatcactatt atattggtca
     2041 tgattaatat gaccaatcgg cgtgtgtttt atatacctct cttatttagt ataagaagat
     2101 cagtactcac ttcttcatta atactaattt ttaacctcta attatcaaca agatctgcgg
     2161 ccgcggccgc aaatttaaat aaaatgaagt gaagttccta tactttctag agaataggaa
     2221 cttctatagt gagtcgaata agggcgacac aaaatttatt ctaaatgcat aataaatact
     2281 gataacatct tatagtttgt attatatttt gtattatcgt tgacatgtat aattttgata
     2341 tcaaaaactg attttccctt tattattttc gagatttatt ttcttaattc tctttaacaa
     2401 actagaaata ttgtatatac aaaaaatcat aaataataga tgaatagttt aattataggt
     2461 gttcatcaat cgaaaaagca acgtatctta tttaaagtgc gttgcttttt tctcatttat
     2521 aaggttaaat aattctcata tatcaagcaa agtgacaggc gcccttaaat attctgacaa
     2581 atgctctttc cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt
     2641 tgcggattaa cgattactcg ttatcagaac cgcccagggg gcccgagctt aagactggcc
     2701 gtcgttttac aacacagaaa gagtttgtag aaacgcaaaa aggccatccg tcaggggcct
     2761 tctgcttagt ttgatgcctg gcagttccct actctcgcct tccgcttcct cgctcactga
     2821 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2881 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2941 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3001 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3061 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3121 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3181 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3241 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3301 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3361 gtatgtaggc ggtgctacag agttcttgaa gtggtgggct aactacggct acactagaag
     3421 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3481 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3541 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3601 cgctcagtgg aacgacgcgc gcgtaactca cgttaaggga ttttggtcat gagcttgcgc
     3661 cgtcccgtca agtcagcgta atgctctgct tttaccaatg cttaatcagt gaggcaccta
     3721 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3781 ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac
     3841 gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3901 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3961 taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg
     4021 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4081 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4141 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4201 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4261 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4321 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4381 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4441 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4501 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc
     4561 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4621 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     4681 acattatcgc gagcccattt atacctgaat atggctcata acaccccttg tttgcctggc
     4741 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     4801 gccgatggta gtgtggggac tccccatgcg agagtaggga actgccaggc atcaaataaa
     4861 acgaaaggct cagtcgaaag actgggcctt tcgcccgggc taattatggg gtgtcgccct
     4921 tattcgactc tatagtgaag ttcctattct ctagaaagta taggaacttc tgaagtgggg
     4981 atttaaatgc ggccgcgctg agggtttaat ggcgcgccgc ggccgcccgc ggtgttggaa
     5041 cgagagtaat taatagtgac atgagttgct atggtaacaa tctaatgctt acatcgtata
     5101 ttaatgtaca actcgtatac gtttaagtgt gattgcgcct attgcagaag gaatgttaaa
     5161 cgagaagctc agacaatact gaagctgtgt taaagaccta ttagttgaac atgttatgct
     5221 agcattaagt cctcagcgag ctcgcatgga atgcgtgcga tgagcgacct catgctatac
     5281 ctgagaaagc aacctgacct acaggaaaga gttactcaag aataagaatt ttcgttttaa
     5341 aacctaagag tcactttaaa atttgtatac acttattttt tttataactt atttaataat
     5401 aaaaatcata aatcataaga aattcgctta tttagaagtg tcaacaacgt atctaccaac
     5461 ggaatgcgtg cgatgcacac accatagctt caaaatgttt ctactccttt tttactcttc
     5521 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
     5581 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
     5641 aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
     5701 tcacgtttct ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc
     5761 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
     5821 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
     5881 agttttaatt acaaatctgt catatgagca aaggagaaga acttttcact ggagttgtcc
     5941 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agaggagagg
     6001 gtgaaggtga tgctacaatc ggaaaactca cccttaaatt tatttgcact actggaaaac
     6061 tacctgttcc atggccaaca cttgtcacta ctctgaccta tggtgttcaa tgcttttccc
     6121 gttatccgga tcacatgaaa aggcatgact ttttcaagag tgccatgccc gaaggttatg
     6181 tacaggaacg cactatatct ttcaaagatg acgggaaata caagacgcgt gctgtagtca
     6241 agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtactgat tttaaagaag
     6301 atggaaacat tctcggacac aaactcgagt acaactttaa ctcacacaat gtatacatca
     6361 cggcagacaa acaaaagaat ggaatcaaag ctaacttcac agttcgccac aacgttgaag
     6421 atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     6481 tccttttacc agacaaccat tacctgtcga cacaaactgt cctttcgaaa gatcccaacg
     6541 aaaagcgtga ccacatggtc cttcatgagt acgtaaatgc tgctgggatt acacatggca
     6601 tggatgagct ctacaaatga atcgcgtg
//
