LOCUS       sequence-148963-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2802 is a vector for multiple integrations at sites sharing 
            homology with Ty1Cons1. 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 7420)
  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 7420)
  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: pCfB2802
FEATURES             Location/Qualifiers
     source          1..7420
                     /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(2989..3022)
                     /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(3492..3508)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3607..3624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3778..4366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3858..3877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4490..5281,5282..5350))
                     /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     5113..5132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5351..5442)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5484..5503
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5509..5528
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5735..5768
                     /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(6054..6219)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6140..6164
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        6268..6674
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6696..7409
                     /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(6726..6754)
                     /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 gttacccctg caggacagac
     2761 gtcattagtg ctgaggcatt aattgatcaa aaacggaatg aggaataatc gtaatattag
     2821 tatgtagaaa tatagattcc attttgagga ttcctatatc ctcgaggaga acttctagtg
     2881 tatattctgt atacctaata ttatagcctt tatcaacaat ggaatcccaa caattatcta
     2941 attacccaca aatttctcaa gatctgcggc cgcactcaga cctgaagtga agttcctata
     3001 ctttctagag aataggaact tctatagtga gtcgaataag ggcgacacaa aatttattct
     3061 aaatgcataa taaatactga taacatctta tagtttgtat tatattttgt attatcgttg
     3121 acatgtataa ttttgatatc aaaaactgat tttcccttta ttattttcga gatttatttt
     3181 cttaattctc tttaacaaac tagaaatatt gtatatacaa aaaatcataa ataatagatg
     3241 aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac gtatcttatt taaagtgcgt
     3301 tgcttttttc tcatttataa ggttaaataa ttctcatata tcaagcaaag tgacaggcgc
     3361 ccttaaatat tctgacaaat gctctttccc taaactcccc ccataaaaaa acccgccgaa
     3421 gcgggttttt acgttatttg cggattaacg attactcgtt atcagaaccg cccagggggc
     3481 ccgagcttaa gactggccgt cgttttacaa cacagaaaga gtttgtagaa acgcaaaaag
     3541 gccatccgtc aggggccttc tgcttagttt gatgcctggc agttccctac tctcgccttc
     3601 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3661 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3721 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3781 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3841 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3901 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3961 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4021 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4081 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4141 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtgggctaa
     4201 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4261 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4321 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4381 cttttctacg gggtctgacg ctcagtggaa cgacgcgcgc gtaactcacg ttaagggatt
     4441 ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat gctctgcttt taccaatgct
     4501 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4561 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agcgctgcga
     4621 tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac cagccagccg
     4681 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4741 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4801 tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4861 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4921 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4981 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5041 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5101 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5161 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5221 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5281 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5341 gaatactcat attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc agtgttacaa
     5461 ccaattaacc aattctgaac attatcgcga gcccatttat acctgaatat ggctcataac
     5521 accccttgtt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     5581 gaagtgaaac gccgtagcgc cgatggtagt gtggggactc cccatgcgag agtagggaac
     5641 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gcccgggcta
     5701 attatggggt gtcgccctta ttcgactcta tagtgaagtt cctattctct agaaagtata
     5761 ggaacttctg aagtggggat ttaaagtcgg tgtccgcgct gagggtttaa tggcgcgccg
     5821 cggccgcccg cggtgttgga ataaaaatcc actatcgtct atcaactaat agttatatta
     5881 tcaatatatt atcatatacg gtgttaagat gatgacataa gttatgagaa gctgtcatcg
     5941 aagttagagg aagctgaagt gcaaggattg ataatgtaat aggatcaatg aatataaaca
     6001 tatgctagca ttaagtcctc agcgagcacg ctgcttcatg gaatgcgtgc gatgagcgac
     6061 ctcatgctat acctgagaaa gcaacctgac ctacaggaaa gagttactca agaataagaa
     6121 ttttcgtttt aaaacctaag agtcacttta aaatttgtat acacttattt tttttataac
     6181 ttatttaata ataaaaatca taaatcataa gaaattcgct tatttagaag tgtcaacaac
     6241 gtatctacca acggaatgcg tgcgatgcac acaccatagc ttcaaaatgt ttctactcct
     6301 tttttactct tccagatttt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca
     6361 agcacagcat actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact
     6421 aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa
     6481 taaaaatttt tatcacgttt ctttttcttg aaaatttttt tttttgattt ttttctcttt
     6541 cgatgacctc ccattgatat ttaagttaat aaacggtctt caatttctca agtttcagtt
     6601 tcatttttct tgttctatta caactttttt tacttcttgc tcattagaaa gaaagcatag
     6661 caatctaatc taagttttaa ttacaaatct gtcatatgag caaaggagaa gaacttttca
     6721 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     6781 tcagaggaga gggtgaaggt gatgctacaa tcggaaaact cacccttaaa tttatttgca
     6841 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactctgacc tatggtgttc
     6901 aatgcttttc ccgttatccg gatcacatga aaaggcatga ctttttcaag agtgccatgc
     6961 ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggaaa tacaagacgc
     7021 gtgctgtagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtactg
     7081 attttaaaga agatggaaac attctcggac acaaactcga gtacaacttt aactcacaca
     7141 atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc acagttcgcc
     7201 acaacgttga agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg
     7261 gcgatggccc tgtcctttta ccagacaacc attacctgtc gacacaaact gtcctttcga
     7321 aagatcccaa cgaaaagcgt gaccacatgg tccttcatga gtacgtaaat gctgctggga
     7381 ttacacatgg catggatgag ctctacaaat gaatcgcgtg
//
