LOCUS       sequence-148926-        6604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2795 is a vector for multiple integrations at sites sharing 
            homology with Ty1Cons1. 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 6604)
  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 6604)
  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: pCfB2795
FEATURES             Location/Qualifiers
     source          1..6604
                     /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(2173..2206)
                     /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(2676..2692)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2791..2808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2962..3550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3042..3061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3674..4465,4466..4534))
                     /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     4297..4316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4535..4626)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4668..4687
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4693..4712
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4919..4952
                     /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(5238..5403)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5324..5348
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5452..5858
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5880..6593
                     /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(5910..5938)
                     /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 tcaaaaacgg
     1981 aatgaggaat aatcgtaata ttagtatgta gaaatataga ttccattttg aggattccta
     2041 tatcctcgag gagaacttct agtgtatatt ctgtatacct aatattatag cctttatcaa
     2101 caatggaatc ccaacaatta tctaattacc cacaaatttc tcaagatctg cggccgcact
     2161 cagacctgaa gtgaagttcc tatactttct agagaatagg aacttctata gtgagtcgaa
     2221 taagggcgac acaaaattta ttctaaatgc ataataaata ctgataacat cttatagttt
     2281 gtattatatt ttgtattatc gttgacatgt ataattttga tatcaaaaac tgattttccc
     2341 tttattattt tcgagattta ttttcttaat tctctttaac aaactagaaa tattgtatat
     2401 acaaaaaatc ataaataata gatgaatagt ttaattatag gtgttcatca atcgaaaaag
     2461 caacgtatct tatttaaagt gcgttgcttt tttctcattt ataaggttaa ataattctca
     2521 tatatcaagc aaagtgacag gcgcccttaa atattctgac aaatgctctt tccctaaact
     2581 ccccccataa aaaaacccgc cgaagcgggt ttttacgtta tttgcggatt aacgattact
     2641 cgttatcaga accgcccagg gggcccgagc ttaagactgg ccgtcgtttt acaacacaga
     2701 aagagtttgt agaaacgcaa aaaggccatc cgtcaggggc cttctgctta gtttgatgcc
     2761 tggcagttcc ctactctcgc cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2821 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2881 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3001 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3061 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3121 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3361 agagttcttg aagtggtggg ctaactacgg ctacactaga agaacagtat ttggtatctg
     3421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3481 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgacgc
     3601 gcgcgtaact cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg
     3661 taatgctctg cttttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3721 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3781 taccatctgg ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt
     3841 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3901 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3961 atagtttgcg caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4021 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4081 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4141 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4201 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4261 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4321 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4381 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4441 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4501 gaataagggc gacacggaaa tgttgaatac tcatattctt cctttttcaa tattattgaa
     4561 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4621 aacaaatagg ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat
     4681 ttatacctga atatggctca taacacccct tgtttgcctg gcggcagtag cgcggtggtc
     4741 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     4801 actccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
     4861 agactgggcc tttcgcccgg gctaattatg gggtgtcgcc cttattcgac tctatagtga
     4921 agttcctatt ctctagaaag tataggaact tctgaagtgg ggatttaaag tcggtgtccg
     4981 cgctgagggt ttaatggcgc gccgcggccg cccgcggtgt tggaataaaa atccactatc
     5041 gtctatcaac taatagttat attatcaata tattatcata tacggtgtta agatgatgac
     5101 ataagttatg agaagctgtc atcgaagtta gaggaagctg aagtgcaagg attgataatg
     5161 taataggatc aatgaatata aacatatgct agcattaagt cctcagcgag cacgctgctt
     5221 catggaatgc gtgcgatgag cgacctcatg ctatacctga gaaagcaacc tgacctacag
     5281 gaaagagtta ctcaagaata agaattttcg ttttaaaacc taagagtcac tttaaaattt
     5341 gtatacactt atttttttta taacttattt aataataaaa atcataaatc ataagaaatt
     5401 cgcttattta gaagtgtcaa caacgtatct accaacggaa tgcgtgcgat gcacacacca
     5461 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
     5521 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
     5581 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
     5641 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     5701 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
     5761 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
     5821 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa atctgtcata
     5881 tgagcaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     5941 atgttaatgg gcacaaattt tctgtcagag gagagggtga aggtgatgct acaatcggaa
     6001 aactcaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
     6061 tcactactct gacctatggt gttcaatgct tttcccgtta tccggatcac atgaaaaggc
     6121 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact atatctttca
     6181 aagatgacgg gaaatacaag acgcgtgctg tagtcaagtt tgaaggtgat acccttgtta
     6241 atcgtatcga gttaaagggt actgatttta aagaagatgg aaacattctc ggacacaaac
     6301 tcgagtacaa ctttaactca cacaatgtat acatcacggc agacaaacaa aagaatggaa
     6361 tcaaagctaa cttcacagtt cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc
     6421 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
     6481 tgtcgacaca aactgtcctt tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc
     6541 atgagtacgt aaatgctgct gggattacac atggcatgga tgagctctac aaatgaatcg
     6601 cgtg
//
