LOCUS       sequence-148880-        6260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2804 is a vector for multiple integrations at sites sharing 
            homology with Ty2Cons. The selective marker is Kl.LEU2. Additionally
            it contains a USER cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6260)
  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 6260)
  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: pCfB2804
FEATURES             Location/Qualifiers
     source          1..6260
                     /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    2607..2640
                     /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(2683..2702)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2684..2702)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2712..2728)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2721..2743)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(2996..3029)
                     /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(3499..3515)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3614..3631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3785..4373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3865..3884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4497..5288,5289..5357))
                     /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     5120..5139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5358..5449)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5491..5510
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5516..5535
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5742..5775
                     /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(6056..6221)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6142..6166
                     /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 aagcagccaa
      481 gatttccttg acagccttgg cgatagcatc gccaacctca gtggtagagt tagaaccacc
      541 aaggtcaccg gttctgacac ctgcatccaa gacatttcta acagcttctt caagagccct
      601 accttcttca accaaatcca aggataactt caacatcata gctgcagata agatggtagc
      661 aattgggtta accttgtttg ctggtaaatc tggggcagaa ccatgacatg gttcgtacaa
      721 accgaatgcc ttgttagtgt caggtaggga agctagagat gcagaaggta ataaacccaa
      781 agaacctgga ataacagagg cttcatcgga gataatatca ccaaacatgt tgttggtaat
      841 aacaacaccg tttagcttag ttggtgattt aaccaaaatc atagcagcag agtcgatcaa
      901 ttggtgctga acagttaatt gtgggaactc agtcttgatg gtttcttcaa cagtctttct
      961 ccacaatctg gaagaggcaa gcacgttagc cttgtcaagt gaccagattg gtaatggtgg
     1021 gttttgttgc aatgccaaga aagcagccat tcttgtaatt ctttgaactt caggaacact
     1081 gtatttctca gagtcccaag caactccgtc accttcatct tcttttcttt caccaaagta
     1141 gataccacca accaattctc taacgacgac gaaatcggta ccctttgcat attcaggctt
     1201 caaaggagaa agatctagta aagaatcaga agcaaagtta catggtctta agttggcgta
     1261 tagacccaat tcctttctga tcttcaatag accttgttct ggtctaactg cgcccgtacc
     1321 ccattttgga ccaccaacag cacctagtaa gacagcatcg gctttcttag aggcttctag
     1381 agcttcatct ggtaaaggag tgccagtggc atcgatggca gcacccccga tcaagtgatg
     1441 ttggaaattg aacttaattt ctggacggac ttcagcaatg gcattcaaga ccttaatagc
     1501 ttcgtcagta acttctttac cgacgtgatc acccggtagg acaacgatat tcttagacat
     1561 ggtataatct gtgtagtgtg ggatactttt tacttcttca aataggtatc aacaataaaa
     1621 ataaatcgaa gcaaatgtag gaatgcgtta aagcagatgt acttacttag agtacataga
     1681 tatatttata taattcaata tataaaagta tatgaagcat ctgatgttga acctgtcatg
     1741 actctaaact ggaatgggta gttatgcttc tgaaggtttt ccgttagaaa atgagtgcag
     1801 ggatcaggcc cggaaccggt tttagcctga aaggaaaaga aaaaaatgcg gtgattacta
     1861 gcacgtgact gcgctgaatt ggaatccacc acaagtacag cgaatggcaa gagggaaggg
     1921 gaatatattg tgcctggcgg tagtctgcta tatcaaatgt taaacataac taaatgcagt
     1981 agtgaataga gtaatgtatt tctacatatg tgggtagggc caatgggagc ccgatgtcta
     2041 ttgtacagat attttccttt tatagttaat caattcaact tggaaatcat tgagtcgata
     2101 tctacttgga taactggagc cgtggcagcg tttgaaattt gacgagtctc cttgttttgt
     2161 tcgcgcattt tcagtttatt caaacgggag atggcaccga atccactgac ttctctgttc
     2221 aatttgaatg gtaataggaa gaaagtacat ttcgagaaat tttcgcccaa cgtcgaaatc
     2281 tcatggatga tatcctcgat acaaatgaca ccttcatcac ctaatttctc ttcaacgata
     2341 ttgttgtcat tcaagattat ctcttttggt tctttatctt ctggcctttg ccacattatt
     2401 ctagatctct tttggattaa agaacggata gatgcaagtg aaggttgtcc gataacaata
     2461 tatggtgata caagtttcaa caatgggtac acgtccttgg taagtttgat gaaaacacca
     2521 gtgttcaact caacaagtct caaaagagat aacactttgt aagcctttgc tgggatcttc
     2581 acagcgagct ctcgagaacc cttaatataa cttcgtataa tgtatgctat acgaagttat
     2641 taggtgatat cagatccact agtggcctat gcacccaatt cgccctatag tgagtcgtat
     2701 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2761 cctgcaggac agacgtcatt agtgctgagg cattaattga tcaatataaa atgatgataa
     2821 taatatttat agaattgtgt agaattgcag attccctttt atggattcct aaatcctgag
     2881 gagaacttct agtatattct acatacctaa tattattgcc ttattaaaaa tggaatccca
     2941 acaattacat caaaatccac attctcagat ctgcggccgc actcagacct gaagtgaagt
     3001 tcctatactt tctagagaat aggaacttct atagtgagtc gaataagggc gacacaaaat
     3061 ttattctaaa tgcataataa atactgataa catcttatag tttgtattat attttgtatt
     3121 atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta ttttcgagat
     3181 ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa atcataaata
     3241 atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta tcttatttaa
     3301 agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca agcaaagtga
     3361 caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca taaaaaaacc
     3421 cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc agaaccgccc
     3481 agggggcccg agcttaagac tggccgtcgt tttacaacac agaaagagtt tgtagaaacg
     3541 caaaaaggcc atccgtcagg ggccttctgc ttagtttgat gcctggcagt tccctactct
     3601 cgccttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3661 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3721 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3781 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3841 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3901 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3961 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4021 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4081 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4141 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4201 gggctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4261 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4321 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4381 ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgcgcgta actcacgtta
     4441 agggattttg gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct ctgcttttac
     4501 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4561 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc
     4621 gctgcgatga taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag
     4681 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4741 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4801 gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4861 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4921 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4981 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5041 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5101 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5161 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5221 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5281 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5341 aaatgttgaa tactcatatt cttccttttt caatattatt gaagcattta tcagggttat
     5401 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt
     5461 gttacaacca attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc
     5521 tcataacacc ccttgtttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     5581 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggactcccc atgcgagagt
     5641 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgcc
     5701 cgggctaatt atggggtgtc gcccttattc gactctatag tgaagttcct attctctaga
     5761 aagtatagga acttctgaag tggggattta aagtcggtgt ccgcgctgag ggtttaatgg
     5821 cgcgccgcgg ccgcccgcgg tgttggaata aaaatcaact atcatctact aactagtatt
     5881 tacgttacta gtatattatc atatacggtg ttagaagatg acgcaaatga tgagaaatag
     5941 tcatctaaat tagtggaagc tgaaacgcaa ggattgataa tgtaatagga tcaatgaata
     6001 ttaacgctag cattaagtcc tcagcgagca cgctgcttca tggaatgcgt gcgatgagcg
     6061 acctcatgct atacctgaga aagcaacctg acctacagga aagagttact caagaataag
     6121 aattttcgtt ttaaaaccta agagtcactt taaaatttgt atacacttat tttttttata
     6181 acttatttaa taataaaaat cataaatcat aagaaattcg cttatttaga agtgtcaaca
     6241 acgtatctac caacggaatg
//
