LOCUS       sequence-148987-        7408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2800 is a vector for multiple integrations at sites sharing 
            homology with Ty2Cons. 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 7408)
  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 7408)
  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: pCfB2800
FEATURES             Location/Qualifiers
     source          1..7408
                     /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(2982..3015)
                     /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(3485..3501)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3600..3617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3771..4359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3851..3870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4483..5274,5275..5343))
                     /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     5106..5125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5344..5435)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5477..5496
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5502..5521
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5728..5761
                     /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(6042..6207)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6128..6152
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        6256..6662
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6684..7397
                     /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(6714..6742)
                     /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 tataaaatga tgataataat atttatagaa
     2821 ttgtgtagaa ttgcagattc ccttttatgg attcctaaat cctgaggaga acttctagta
     2881 tattctacat acctaatatt attgccttat taaaaatgga atcccaacaa ttacatcaaa
     2941 atccacattc tcagatctgc ggccgcactc agacctgaag tgaagttcct atactttcta
     3001 gagaatagga acttctatag tgagtcgaat aagggcgaca caaaatttat tctaaatgca
     3061 taataaatac tgataacatc ttatagtttg tattatattt tgtattatcg ttgacatgta
     3121 taattttgat atcaaaaact gattttccct ttattatttt cgagatttat tttcttaatt
     3181 ctctttaaca aactagaaat attgtatata caaaaaatca taaataatag atgaatagtt
     3241 taattatagg tgttcatcaa tcgaaaaagc aacgtatctt atttaaagtg cgttgctttt
     3301 ttctcattta taaggttaaa taattctcat atatcaagca aagtgacagg cgcccttaaa
     3361 tattctgaca aatgctcttt ccctaaactc cccccataaa aaaacccgcc gaagcgggtt
     3421 tttacgttat ttgcggatta acgattactc gttatcagaa ccgcccaggg ggcccgagct
     3481 taagactggc cgtcgtttta caacacagaa agagtttgta gaaacgcaaa aaggccatcc
     3541 gtcaggggcc ttctgcttag tttgatgcct ggcagttccc tactctcgcc ttccgcttcc
     3601 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3661 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3721 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3781 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3841 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3901 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3961 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4021 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4081 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4141 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtgggc taactacggc
     4201 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4261 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4321 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4381 acggggtctg acgctcagtg gaacgacgcg cgcgtaactc acgttaaggg attttggtca
     4441 tgagcttgcg ccgtcccgtc aagtcagcgt aatgctctgc ttttaccaat gcttaatcag
     4501 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4561 cgtgtagata actacgatac gggagggctt accatctggc cccagcgctg cgatgatacc
     4621 gcgagaacca cgctcaccgg ctccggattt atcagcaata aaccagccag ccggaagggc
     4681 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4741 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccatcgctac
     4801 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4861 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4921 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4981 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5041 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5101 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5161 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5221 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5281 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5341 catattcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5401 atacatattt gaatgtattt agaaaaataa acaaataggg gtcagtgtta caaccaatta
     5461 accaattctg aacattatcg cgagcccatt tatacctgaa tatggctcat aacacccctt
     5521 gtttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     5581 aacgccgtag cgccgatggt agtgtgggga ctccccatgc gagagtaggg aactgccagg
     5641 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgcccggg ctaattatgg
     5701 ggtgtcgccc ttattcgact ctatagtgaa gttcctattc tctagaaagt ataggaactt
     5761 ctgaagtggg gatttaaagt cggtgtccgc gctgagggtt taatggcgcg ccgcggccgc
     5821 ccgcggtgtt ggaataaaaa tcaactatca tctactaact agtatttacg ttactagtat
     5881 attatcatat acggtgttag aagatgacgc aaatgatgag aaatagtcat ctaaattagt
     5941 ggaagctgaa acgcaaggat tgataatgta ataggatcaa tgaatattaa cgctagcatt
     6001 aagtcctcag cgagcacgct gcttcatgga atgcgtgcga tgagcgacct catgctatac
     6061 ctgagaaagc aacctgacct acaggaaaga gttactcaag aataagaatt ttcgttttaa
     6121 aacctaagag tcactttaaa atttgtatac acttattttt tttataactt atttaataat
     6181 aaaaatcata aatcataaga aattcgctta tttagaagtg tcaacaacgt atctaccaac
     6241 ggaatgcgtg cgatgcacac accatagctt caaaatgttt ctactccttt tttactcttc
     6301 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
     6361 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
     6421 aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
     6481 tcacgtttct ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc
     6541 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
     6601 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
     6661 agttttaatt acaaatctgt catatgagca aaggagaaga acttttcact ggagttgtcc
     6721 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agaggagagg
     6781 gtgaaggtga tgctacaatc ggaaaactca cccttaaatt tatttgcact actggaaaac
     6841 tacctgttcc atggccaaca cttgtcacta ctctgaccta tggtgttcaa tgcttttccc
     6901 gttatccgga tcacatgaaa aggcatgact ttttcaagag tgccatgccc gaaggttatg
     6961 tacaggaacg cactatatct ttcaaagatg acgggaaata caagacgcgt gctgtagtca
     7021 agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtactgat tttaaagaag
     7081 atggaaacat tctcggacac aaactcgagt acaactttaa ctcacacaat gtatacatca
     7141 cggcagacaa acaaaagaat ggaatcaaag ctaacttcac agttcgccac aacgttgaag
     7201 atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     7261 tccttttacc agacaaccat tacctgtcga cacaaactgt cctttcgaaa gatcccaacg
     7321 aaaagcgtga ccacatggtc cttcatgagt acgtaaatgc tgctgggatt acacatggca
     7381 tggatgagct ctacaaatga atcgcgtg
//
