LOCUS       sequence-148848-        5456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2989 is a vector for multiple integrations at sites sharing 
            homology with Ty1Cons1. The selective marker is Kl.URA3. 
            Additionally it contains a USER cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5456)
  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 5456)
  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: pCfB2989
FEATURES             Location/Qualifiers
     source          1..5456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      20..209
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     protein_bind    257..290
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1791..1824
                     /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(1867..1886)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1868..1886)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1896..1912)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1905..1927)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(2187..2220)
                     /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(2690..2706)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2805..2822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2976..3564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3056..3075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3688..4479,4480..4548))
                     /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     4311..4330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4549..4640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4682..4701
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4707..4726
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4933..4966
                     /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(5252..5417)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5338..5362
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 cgtgcgatcg cgtgcattca tccgctctaa ccgaaaagga aggagttaga caacctgaag
       61 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
      121 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
      181 tgcttgagaa ggttttggga cgctcgaaga tcgcgtcagc tgaagcttcg tacgctgcag
      241 gtcgacaacc cttaatataa cttcgtataa tgtatgctat acgaagttat taggtctaga
      301 gatcccaata caacagatca cgtgatcttt ttgtaagatg aagttgaagt agtgttgcac
      361 cgtgccaatg caggtggcta ttagattaaa tatgtgattt gttctattaa gtttcctgta
      421 taattacaaa tgaataacga aatgagacaa agaagagaac caatttttac aagcatgggg
      481 agcgctgatt ctcttttggt acgcttccca tccagcattt ctgtatcttt caccttcaac
      541 cttaggatct ctacccttgg cgaaaagtcc tctgccaaca atgatgatat ctgatccacc
      601 acttacaact tcgtcgacgg ttctgtactg ctgacccaat gcatcgcctt tgtcgtctaa
      661 acctacacct ggggtcatga ttagccaatc aaacccttct tctcttcctc ccatatcgtt
      721 ctgagcaatg aacccaataa cgaaatcttt atcactcttt gcaatatcaa cggtaccctt
      781 agtatattca ccgtgtgcta gagaaccctt ggaagataat tcagcaagca tcaataatcc
      841 ccttggttct ttggtgacct cttgcgcacc ttgtttcaag ccagcaacaa taccagcacc
      901 agtaaccccg tgggcgttgg tgatatcaga ccattctgcg atacggtaaa cgcccgatgt
      961 atattgtaat ttgactgtgt taccgatatc ggcgaatttt ctgtcctcaa atatcaagaa
     1021 cttgtatttc tctgccaatg ctttcaatgg aacgacagta ccctcataac tgaaatcatc
     1081 caagatatca acgtgtgttt tcaaaaggca aatgtatgga cccaacgttt caacaagttt
     1141 caatagctca tcagtcgaac gaacgtcaag agaagcacac aaattggttt tcttttcatc
     1201 cattaaacgt aaaagtttcg atgcaaccgg acttgcatga gtctcagctc tactggtata
     1261 tgattttgtg gacatggtgc aactaattga cgggagtgta ttgacgctgg cgtactggct
     1321 ttcacaaaat ggcccaatca caaccacatc ttagatagtt gaaatgactt tagataacat
     1381 caattgagat gagcttaatc atgtcaaagc taaaagtgtc accatgaacg acaattctta
     1441 agcaaatcac gtgatataga tccacgaata accaccattt gatgctcgag gcaagtaatg
     1501 tgtgtaaaaa aatgcgttac caccatccaa tgcagaccga tcttctaccc agaatcacat
     1561 atatttatgt accgagtacc ttttttctat cttccaattg cttctcccat atgattgtct
     1621 ccgtaagctc gaaatttcta agttggattt taatcttcac gcaggatgac agttcgatga
     1681 gcttctgagg agtgtttaga acataatcag tttatccatg gtctatctct tcttgtcgct
     1741 ttttctcctc gatagaacct aaataaaacg agctctcgag aacccttaat ataacttcgt
     1801 ataatgtatg ctatacgaag ttattaggtg atatcagatc cactagtggc ctatgcaccc
     1861 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     1921 gactgggaaa accctggcgt tacccctgca ggacagacgt cattagtgct gaggcattaa
     1981 ttgatcaaaa acggaatgag gaataatcgt aatattagta tgtagaaata tagattccat
     2041 tttgaggatt cctatatcct cgaggagaac ttctagtgta tattctgtat acctaatatt
     2101 atagccttta tcaacaatgg aatcccaaca attatctaat tacccacaaa tttctcaaga
     2161 tctgcggccg cactcagacc tgaagtgaag ttcctatact ttctagagaa taggaacttc
     2221 tatagtgagt cgaataaggg cgacacaaaa tttattctaa atgcataata aatactgata
     2281 acatcttata gtttgtatta tattttgtat tatcgttgac atgtataatt ttgatatcaa
     2341 aaactgattt tccctttatt attttcgaga tttattttct taattctctt taacaaacta
     2401 gaaatattgt atatacaaaa aatcataaat aatagatgaa tagtttaatt ataggtgttc
     2461 atcaatcgaa aaagcaacgt atcttattta aagtgcgttg cttttttctc atttataagg
     2521 ttaaataatt ctcatatatc aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc
     2581 tctttcccta aactcccccc ataaaaaaac ccgccgaagc gggtttttac gttatttgcg
     2641 gattaacgat tactcgttat cagaaccgcc cagggggccc gagcttaaga ctggccgtcg
     2701 ttttacaaca cagaaagagt ttgtagaaac gcaaaaaggc catccgtcag gggccttctg
     2761 cttagtttga tgcctggcag ttccctactc tcgccttccg cttcctcgct cactgactcg
     2821 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2881 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2941 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3001 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3061 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3121 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3181 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3241 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3301 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3361 gtaggcggtg ctacagagtt cttgaagtgg tgggctaact acggctacac tagaagaaca
     3421 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3481 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3541 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3601 cagtggaacg acgcgcgcgt aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc
     3661 ccgtcaagtc agcgtaatgc tctgctttta ccaatgctta atcagtgagg cacctatctc
     3721 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3781 gatacgggag ggcttaccat ctggccccag cgctgcgatg ataccgcgag aaccacgctc
     3841 accggctccg gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3901 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3961 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc
     4021 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4081 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4141 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4201 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4261 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4321 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4381 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4441 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4501 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatat tcttcctttt
     4561 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4621 tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa ttctgaacat
     4681 tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgtttg cctggcggca
     4741 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     4801 atggtagtgt ggggactccc catgcgagag tagggaactg ccaggcatca aataaaacga
     4861 aaggctcagt cgaaagactg ggcctttcgc ccgggctaat tatggggtgt cgcccttatt
     4921 cgactctata gtgaagttcc tattctctag aaagtatagg aacttctgaa gtggggattt
     4981 aaagtcggtg tccgcgctga gggtttaatg gcgcgccgcg gccgcccgcg gtgttggaat
     5041 aaaaatccac tatcgtctat caactaatag ttatattatc aatatattat catatacggt
     5101 gttaagatga tgacataagt tatgagaagc tgtcatcgaa gttagaggaa gctgaagtgc
     5161 aaggattgat aatgtaatag gatcaatgaa tataaacata tgctagcatt aagtcctcag
     5221 cgagcacgct gcttcatgga atgcgtgcga tgagcgacct catgctatac ctgagaaagc
     5281 aacctgacct acaggaaaga gttactcaag aataagaatt ttcgttttaa aacctaagag
     5341 tcactttaaa atttgtatac acttattttt tttataactt atttaataat aaaaatcata
     5401 aatcataaga aattcgctta tttagaagtg tcaacaacgt atctaccaac ggaatg
//
