LOCUS       sequence-149000-        7421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2799 is a vector for multiple integrations at sites sharing 
            homology with Ty3Cons. 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 7421)
  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 7421)
  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: pCfB2799
FEATURES             Location/Qualifiers
     source          1..7421
                     /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(2990..3023)
                     /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(3493..3509)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3608..3625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3779..4367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3859..3878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4491..5282,5283..5351))
                     /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     5114..5133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5352..5443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5485..5504
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5510..5529
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5736..5769
                     /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(6055..6220)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6141..6165
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        6269..6675
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6697..7410
                     /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(6727..6755)
                     /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 aattgatcag attccgcgct tccaccactt agtatgattc
     2821 atattttata taatatataa gataagtaac attccgtgaa ttaatctgat aaactgtttt
     2881 gacaactggt tacttcccta agactgttta tattaggatt gtcaagacac tccggtatta
     2941 ctcgagcccg taatacaaca agatctgcgg ccgcactcag acctgaagtg aagttcctat
     3001 actttctaga gaataggaac ttctatagtg agtcgaataa gggcgacaca aaatttattc
     3061 taaatgcata ataaatactg ataacatctt atagtttgta ttatattttg tattatcgtt
     3121 gacatgtata attttgatat caaaaactga ttttcccttt attattttcg agatttattt
     3181 tcttaattct ctttaacaaa ctagaaatat tgtatataca aaaaatcata aataatagat
     3241 gaatagttta attataggtg ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg
     3301 ttgctttttt ctcatttata aggttaaata attctcatat atcaagcaaa gtgacaggcg
     3361 cccttaaata ttctgacaaa tgctctttcc ctaaactccc cccataaaaa aacccgccga
     3421 agcgggtttt tacgttattt gcggattaac gattactcgt tatcagaacc gcccaggggg
     3481 cccgagctta agactggccg tcgttttaca acacagaaag agtttgtaga aacgcaaaaa
     3541 ggccatccgt caggggcctt ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt
     3601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3961 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtgggcta
     4201 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4381 tcttttctac ggggtctgac gctcagtgga acgacgcgcg cgtaactcac gttaagggat
     4441 tttggtcatg agcttgcgcc gtcccgtcaa gtcagcgtaa tgctctgctt ttaccaatgc
     4501 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4561 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagcgctgcg
     4621 atgataccgc gagaaccacg ctcaccggct ccggatttat cagcaataaa ccagccagcc
     4681 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4741 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4801 atcgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4861 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4921 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4981 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5041 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5101 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5161 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5221 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5281 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5341 tgaatactca tattcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5401 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt cagtgttaca
     5461 accaattaac caattctgaa cattatcgcg agcccattta tacctgaata tggctcataa
     5521 caccccttgt ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     5581 agaagtgaaa cgccgtagcg ccgatggtag tgtggggact ccccatgcga gagtagggaa
     5641 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgcccgggct
     5701 aattatgggg tgtcgccctt attcgactct atagtgaagt tcctattctc tagaaagtat
     5761 aggaacttct gaagtgggga tttaaagtcg gtgtccgcgc tgagggttta atggcgcgcc
     5821 gcggccgccc gcggtgttgt atctcaaaat gagatatgtc agtatgacaa tacgtcatcc
     5881 tgaacgttca taaaacacat atgaaacaac cttataacaa aacgaacaac atgagacaaa
     5941 acccgtcctt ccctagctga actacccaaa agtataaatg cctgaacaat tagtttagat
     6001 ccgagctagc attaagtcct cagcgagcac gctgcttcat ggaatgcgtg cgatgagcga
     6061 cctcatgcta tacctgagaa agcaacctga cctacaggaa agagttactc aagaataaga
     6121 attttcgttt taaaacctaa gagtcacttt aaaatttgta tacacttatt ttttttataa
     6181 cttatttaat aataaaaatc ataaatcata agaaattcgc ttatttagaa gtgtcaacaa
     6241 cgtatctacc aacggaatgc gtgcgatgca cacaccatag cttcaaaatg tttctactcc
     6301 ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc
     6361 aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac
     6421 taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca
     6481 ataaaaattt ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt
     6541 tcgatgacct cccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt
     6601 ttcatttttc ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata
     6661 gcaatctaat ctaagtttta attacaaatc tgtcatatga gcaaaggaga agaacttttc
     6721 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
     6781 gtcagaggag agggtgaagg tgatgctaca atcggaaaac tcacccttaa atttatttgc
     6841 actactggaa aactacctgt tccatggcca acacttgtca ctactctgac ctatggtgtt
     6901 caatgctttt cccgttatcc ggatcacatg aaaaggcatg actttttcaa gagtgccatg
     6961 cccgaaggtt atgtacagga acgcactata tctttcaaag atgacgggaa atacaagacg
     7021 cgtgctgtag tcaagtttga aggtgatacc cttgttaatc gtatcgagtt aaagggtact
     7081 gattttaaag aagatggaaa cattctcgga cacaaactcg agtacaactt taactcacac
     7141 aatgtataca tcacggcaga caaacaaaag aatggaatca aagctaactt cacagttcgc
     7201 cacaacgttg aagatggttc cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     7261 ggcgatggcc ctgtcctttt accagacaac cattacctgt cgacacaaac tgtcctttcg
     7321 aaagatccca acgaaaagcg tgaccacatg gtccttcatg agtacgtaaa tgctgctggg
     7381 attacacatg gcatggatga gctctacaaa tgaatcgcgt g
//
