LOCUS       sequence-148909-        6296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2803 is a vector for multiple integrations at sites sharing 
            homology with Ty4Cons. 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 6296)
  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 6296)
  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: pCfB2803
FEATURES             Location/Qualifiers
     source          1..6296
                     /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(3021..3054)
                     /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(3524..3540)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3639..3656)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3810..4398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3890..3909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4522..5313,5314..5382))
                     /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     5145..5164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5383..5474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5516..5535
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5541..5560
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5767..5800
                     /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(6092..6257)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6178..6202
                     /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 tagtgctgag gcattaattg atcaggtagg tacatatatg aggaatatga
     2821 gtcgtcacat caatgtatag taactaccgg aatcactatt atattggtca tgattaatat
     2881 gaccaatcgg cgtgtgtttt atatacctct cttatttagt ataagaagat cagtactcac
     2941 ttcttcatta atactaattt ttaacctcta attatcaaca agatctgcgg ccgcggccgc
     3001 aaatttaaat aaaatgaagt gaagttccta tactttctag agaataggaa cttctatagt
     3061 gagtcgaata agggcgacac aaaatttatt ctaaatgcat aataaatact gataacatct
     3121 tatagtttgt attatatttt gtattatcgt tgacatgtat aattttgata tcaaaaactg
     3181 attttccctt tattattttc gagatttatt ttcttaattc tctttaacaa actagaaata
     3241 ttgtatatac aaaaaatcat aaataataga tgaatagttt aattataggt gttcatcaat
     3301 cgaaaaagca acgtatctta tttaaagtgc gttgcttttt tctcatttat aaggttaaat
     3361 aattctcata tatcaagcaa agtgacaggc gcccttaaat attctgacaa atgctctttc
     3421 cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt tgcggattaa
     3481 cgattactcg ttatcagaac cgcccagggg gcccgagctt aagactggcc gtcgttttac
     3541 aacacagaaa gagtttgtag aaacgcaaaa aggccatccg tcaggggcct tctgcttagt
     3601 ttgatgcctg gcagttccct actctcgcct tccgcttcct cgctcactga ctcgctgcgc
     3661 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3721 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3781 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3841 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3901 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3961 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4021 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4081 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4141 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4201 ggtgctacag agttcttgaa gtggtgggct aactacggct acactagaag aacagtattt
     4261 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4321 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4381 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4441 aacgacgcgc gcgtaactca cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca
     4501 agtcagcgta atgctctgct tttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4561 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4621 ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac gctcaccggc
     4681 tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4741 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4801 gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg tgtcacgctc
     4861 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4921 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4981 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5041 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5101 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5161 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5221 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5281 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5341 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc tttttcaata
     5401 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5461 gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga acattatcgc
     5521 gagcccattt atacctgaat atggctcata acaccccttg tttgcctggc ggcagtagcg
     5581 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     5641 gtgtggggac tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     5701 cagtcgaaag actgggcctt tcgcccgggc taattatggg gtgtcgccct tattcgactc
     5761 tatagtgaag ttcctattct ctagaaagta taggaacttc tgaagtgggg atttaaatgc
     5821 ggccgcgctg agggtttaat ggcgcgccgc ggccgcccgc ggtgttggaa cgagagtaat
     5881 taatagtgac atgagttgct atggtaacaa tctaatgctt acatcgtata ttaatgtaca
     5941 actcgtatac gtttaagtgt gattgcgcct attgcagaag gaatgttaaa cgagaagctc
     6001 agacaatact gaagctgtgt taaagaccta ttagttgaac atgttatgct agcattaagt
     6061 cctcagcgag ctcgcatgga atgcgtgcga tgagcgacct catgctatac ctgagaaagc
     6121 aacctgacct acaggaaaga gttactcaag aataagaatt ttcgttttaa aacctaagag
     6181 tcactttaaa atttgtatac acttattttt tttataactt atttaataat aaaaatcata
     6241 aatcataaga aattcgctta tttagaagtg tcaacaacgt atctaccaac ggaatg
//
