LOCUS       sequence-150442-        5453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2988 is a vector for multiple integrations at sites sharing 
            homology with Ty1Cons2. 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 5453)
  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 5453)
  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: pCfB2988
FEATURES             Location/Qualifiers
     source          1..5453
                     /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(2186..2219)
                     /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(2689..2705)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2804..2821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2975..3563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3055..3074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3687..4478,4479..4547))
                     /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     4310..4329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4548..4639)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4681..4700
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4706..4725
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4932..4965
                     /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(5249..5414)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5335..5359
                     /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 ttgatcataa aacggaatga tgaataatat ttatagaatt gtgtagaatt gcagattccc
     2041 ttttatggat tcctaaatcc tcgaggagaa cttctagtat attctgtata cctaatatta
     2101 tagcctttat caacaatgga atcccaacaa ttatctcaaa attcacatat ttctcaagat
     2161 ctgcggccgc actcagacct gaagtgaagt tcctatactt tctagagaat aggaacttct
     2221 atagtgagtc gaataagggc gacacaaaat ttattctaaa tgcataataa atactgataa
     2281 catcttatag tttgtattat attttgtatt atcgttgaca tgtataattt tgatatcaaa
     2341 aactgatttt ccctttatta ttttcgagat ttattttctt aattctcttt aacaaactag
     2401 aaatattgta tatacaaaaa atcataaata atagatgaat agtttaatta taggtgttca
     2461 tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc ttttttctca tttataaggt
     2521 taaataattc tcatatatca agcaaagtga caggcgccct taaatattct gacaaatgct
     2581 ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg
     2641 attaacgatt actcgttatc agaaccgccc agggggcccg agcttaagac tggccgtcgt
     2701 tttacaacac agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc
     2761 ttagtttgat gcctggcagt tccctactct cgccttccgc ttcctcgctc actgactcgc
     2821 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2881 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2941 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3001 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3061 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3121 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3181 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3241 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3301 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3361 taggcggtgc tacagagttc ttgaagtggt gggctaacta cggctacact agaagaacag
     3421 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3481 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3541 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3601 agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagct tgcgccgtcc
     3661 cgtcaagtca gcgtaatgct ctgcttttac caatgcttaa tcagtgaggc acctatctca
     3721 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3781 atacgggagg gcttaccatc tggccccagc gctgcgatga taccgcgaga accacgctca
     3841 ccggctccgg atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3901 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3961 agttcgccag ttaatagttt gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca
     4021 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4081 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4141 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4201 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4261 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4321 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4381 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4441 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4501 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatatt cttccttttt
     4561 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4621 atttagaaaa ataaacaaat aggggtcagt gttacaacca attaaccaat tctgaacatt
     4681 atcgcgagcc catttatacc tgaatatggc tcataacacc ccttgtttgc ctggcggcag
     4741 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     4801 tggtagtgtg gggactcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     4861 aggctcagtc gaaagactgg gcctttcgcc cgggctaatt atggggtgtc gcccttattc
     4921 gactctatag tgaagttcct attctctaga aagtatagga acttctgaag tggggattta
     4981 aagtcggtgt ccgcgctgag ggtttaatgg cgcgccgcgg ccgcccgcgg tgttggaata
     5041 aaaatcaact atcatctact aactagtatt tacgttacta gtatattatc atatacggtg
     5101 ttagaagatg acgcaaatga tgagaaatag tcatctaaat tagtggaagc tgaaacgcaa
     5161 ggattgataa tgtaatagga tcaatgaata ttaacatagc tagcattaag tcctcagcga
     5221 gcacgctgct tcatggaatg cgtgcgatga gcgacctcat gctatacctg agaaagcaac
     5281 ctgacctaca ggaaagagtt actcaagaat aagaattttc gttttaaaac ctaagagtca
     5341 ctttaaaatt tgtatacact tatttttttt ataacttatt taataataaa aatcataaat
     5401 cataagaaat tcgcttattt agaagtgtca acaacgtatc taccaacgga atg
//
