LOCUS       sequence-148851-        5457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2875 is a vector for multiple integrations at sites sharing 
            homology with Ty3Cons. 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 5457)
  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 5457)
  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: pCfB2875
FEATURES             Location/Qualifiers
     source          1..5457
                     /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(2188..2221)
                     /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(2691..2707)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2806..2823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2977..3565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3057..3076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3689..4480,4481..4549))
                     /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     4312..4331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4550..4641)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4683..4702
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4708..4727
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4934..4967
                     /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(5253..5418)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5339..5363
                     /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 ttgatcagat tccgcgcttc caccacttag tatgattcat attttatata atatataaga
     2041 taagtaacat tccgtgaatt aatctgataa actgttttga caactggtta cttccctaag
     2101 actgtttata ttaggattgt caagacactc cggtattact cgagcccgta atacaacaag
     2161 atctgcggcc gcactcagac ctgaagtgaa gttcctatac tttctagaga ataggaactt
     2221 ctatagtgag tcgaataagg gcgacacaaa atttattcta aatgcataat aaatactgat
     2281 aacatcttat agtttgtatt atattttgta ttatcgttga catgtataat tttgatatca
     2341 aaaactgatt ttccctttat tattttcgag atttattttc ttaattctct ttaacaaact
     2401 agaaatattg tatatacaaa aaatcataaa taatagatga atagtttaat tataggtgtt
     2461 catcaatcga aaaagcaacg tatcttattt aaagtgcgtt gcttttttct catttataag
     2521 gttaaataat tctcatatat caagcaaagt gacaggcgcc cttaaatatt ctgacaaatg
     2581 ctctttccct aaactccccc cataaaaaaa cccgccgaag cgggttttta cgttatttgc
     2641 ggattaacga ttactcgtta tcagaaccgc ccagggggcc cgagcttaag actggccgtc
     2701 gttttacaac acagaaagag tttgtagaaa cgcaaaaagg ccatccgtca ggggccttct
     2761 gcttagtttg atgcctggca gttccctact ctcgccttcc gcttcctcgc tcactgactc
     2821 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2881 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2941 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3001 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3061 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3121 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3181 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3241 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3301 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3361 tgtaggcggt gctacagagt tcttgaagtg gtgggctaac tacggctaca ctagaagaac
     3421 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3481 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3601 tcagtggaac gacgcgcgcg taactcacgt taagggattt tggtcatgag cttgcgccgt
     3661 cccgtcaagt cagcgtaatg ctctgctttt accaatgctt aatcagtgag gcacctatct
     3721 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3781 cgatacggga gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct
     3841 caccggctcc ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3901 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3961 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt
     4021 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4081 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4141 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4201 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4261 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4321 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4381 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4441 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4501 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ttcttccttt
     4561 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4621 gtatttagaa aaataaacaa ataggggtca gtgttacaac caattaacca attctgaaca
     4681 ttatcgcgag cccatttata cctgaatatg gctcataaca ccccttgttt gcctggcggc
     4741 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     4801 gatggtagtg tggggactcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     4861 aaaggctcag tcgaaagact gggcctttcg cccgggctaa ttatggggtg tcgcccttat
     4921 tcgactctat agtgaagttc ctattctcta gaaagtatag gaacttctga agtggggatt
     4981 taaagtcggt gtccgcgctg agggtttaat ggcgcgccgc ggccgcccgc ggtgttgtat
     5041 ctcaaaatga gatatgtcag tatgacaata cgtcatcctg aacgttcata aaacacatat
     5101 gaaacaacct tataacaaaa cgaacaacat gagacaaaac ccgtccttcc ctagctgaac
     5161 tacccaaaag tataaatgcc tgaacaatta gtttagatcc gagctagcat taagtcctca
     5221 gcgagcacgc tgcttcatgg aatgcgtgcg atgagcgacc tcatgctata cctgagaaag
     5281 caacctgacc tacaggaaag agttactcaa gaataagaat tttcgtttta aaacctaaga
     5341 gtcactttaa aatttgtata cacttatttt ttttataact tatttaataa taaaaatcat
     5401 aaatcataag aaattcgctt atttagaagt gtcaacaacg tatctaccaa cggaatg
//
