LOCUS       sequence-148857-        5480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2796 is a vector for multiple integrations at sites sharing 
            homology with Ty4Cons. 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 5480)
  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 5480)
  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: pCfB2796
FEATURES             Location/Qualifiers
     source          1..5480
                     /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(2205..2238)
                     /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(2708..2724)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2823..2840)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2994..3582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3074..3093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3706..4497,4498..4566))
                     /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     4329..4348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4567..4658)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4700..4719
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4725..4744
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4951..4984
                     /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(5276..5441)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5362..5386
                     /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 tacccctaca ggactagtgc tgaggcatta attgatcagg
     1981 taggtacata tatgaggaat atgagtcgtc acatcaatgt atagtaacta ccggaatcac
     2041 tattatattg gtcatgatta atatgaccaa tcggcgtgtg ttttatatac ctctcttatt
     2101 tagtataaga agatcagtac tcacttcttc attaatacta atttttaacc tctaattatc
     2161 aacaagatct gcggccgcgg ccgcaaattt aaataaaatg aagtgaagtt cctatacttt
     2221 ctagagaata ggaacttcta tagtgagtcg aataagggcg acacaaaatt tattctaaat
     2281 gcataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
     2341 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
     2401 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
     2461 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
     2521 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
     2581 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
     2641 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
     2701 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
     2761 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
     2821 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2881 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2941 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3001 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3061 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3121 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3181 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3241 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3301 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3361 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
     3421 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3481 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3541 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3601 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
     3661 tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc tgcttttacc aatgcttaat
     3721 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3781 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagcg ctgcgatgat
     3841 accgcgagaa ccacgctcac cggctccgga tttatcagca ataaaccagc cagccggaag
     3901 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3961 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccatcgc
     4021 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4081 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4141 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4201 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4261 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4321 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4381 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4441 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4501 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4561 actcatattc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4621 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa
     4681 ttaaccaatt ctgaacatta tcgcgagccc atttatacct gaatatggct cataacaccc
     4741 cttgtttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     4801 tgaaacgccg tagcgccgat ggtagtgtgg ggactcccca tgcgagagta gggaactgcc
     4861 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgccc gggctaatta
     4921 tggggtgtcg cccttattcg actctatagt gaagttccta ttctctagaa agtataggaa
     4981 cttctgaagt ggggatttaa atgcggccgc gctgagggtt taatggcgcg ccgcggccgc
     5041 ccgcggtgtt ggaacgagag taattaatag tgacatgagt tgctatggta acaatctaat
     5101 gcttacatcg tatattaatg tacaactcgt atacgtttaa gtgtgattgc gcctattgca
     5161 gaaggaatgt taaacgagaa gctcagacaa tactgaagct gtgttaaaga cctattagtt
     5221 gaacatgtta tgctagcatt aagtcctcag cgagctcgca tggaatgcgt gcgatgagcg
     5281 acctcatgct atacctgaga aagcaacctg acctacagga aagagttact caagaataag
     5341 aattttcgtt ttaaaaccta agagtcactt taaaatttgt atacacttat tttttttata
     5401 acttatttaa taataaaaat cataaatcat aagaaattcg cttatttaga agtgtcaaca
     5461 acgtatctac caacggaatg
//
