LOCUS       sequence-148883-        6273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2990 is a vector for multiple integrations at sites sharing 
            homology with Ty3Cons. 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 6273)
  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 6273)
  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: pCfB2990
FEATURES             Location/Qualifiers
     source          1..6273
                     /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(3004..3037)
                     /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(3507..3523)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3622..3639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3793..4381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3873..3892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4505..5296,5297..5365))
                     /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     5128..5147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5366..5457)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5499..5518
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5524..5543
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5750..5783
                     /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(6069..6234)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6155..6179
                     /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 tcagattccg cgcttccacc
     2821 acttagtatg attcatattt tatataatat ataagataag taacattccg tgaattaatc
     2881 tgataaactg ttttgacaac tggttacttc cctaagactg tttatattag gattgtcaag
     2941 acactccggt attactcgag cccgtaatac aacaagatct gcggccgcac tcagacctga
     3001 agtgaagttc ctatactttc tagagaatag gaacttctat agtgagtcga ataagggcga
     3061 cacaaaattt attctaaatg cataataaat actgataaca tcttatagtt tgtattatat
     3121 tttgtattat cgttgacatg tataattttg atatcaaaaa ctgattttcc ctttattatt
     3181 ttcgagattt attttcttaa ttctctttaa caaactagaa atattgtata tacaaaaaat
     3241 cataaataat agatgaatag tttaattata ggtgttcatc aatcgaaaaa gcaacgtatc
     3301 ttatttaaag tgcgttgctt ttttctcatt tataaggtta aataattctc atatatcaag
     3361 caaagtgaca ggcgccctta aatattctga caaatgctct ttccctaaac tccccccata
     3421 aaaaaacccg ccgaagcggg tttttacgtt atttgcggat taacgattac tcgttatcag
     3481 aaccgcccag ggggcccgag cttaagactg gccgtcgttt tacaacacag aaagagtttg
     3541 tagaaacgca aaaaggccat ccgtcagggg ccttctgctt agtttgatgc ctggcagttc
     3601 cctactctcg ccttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3661 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3721 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3781 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3841 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3901 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3961 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4021 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4081 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4141 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4201 gaagtggtgg gctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4261 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4321 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4381 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgcgcgtaac
     4441 tcacgttaag ggattttggt catgagcttg cgccgtcccg tcaagtcagc gtaatgctct
     4501 gcttttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4561 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4621 gccccagcgc tgcgatgata ccgcgagaac cacgctcacc ggctccggat ttatcagcaa
     4681 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4741 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4801 gcaacgttgt tgccatcgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4861 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4921 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4981 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5041 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5101 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5161 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5221 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5281 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5341 cgacacggaa atgttgaata ctcatattct tcctttttca atattattga agcatttatc
     5401 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5461 gggtcagtgt tacaaccaat taaccaattc tgaacattat cgcgagccca tttatacctg
     5521 aatatggctc ataacacccc ttgtttgcct ggcggcagta gcgcggtggt cccacctgac
     5581 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gactccccat
     5641 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     5701 ctttcgcccg ggctaattat ggggtgtcgc ccttattcga ctctatagtg aagttcctat
     5761 tctctagaaa gtataggaac ttctgaagtg gggatttaaa gtcggtgtcc gcgctgaggg
     5821 tttaatggcg cgccgcggcc gcccgcggtg ttgtatctca aaatgagata tgtcagtatg
     5881 acaatacgtc atcctgaacg ttcataaaac acatatgaaa caaccttata acaaaacgaa
     5941 caacatgaga caaaacccgt ccttccctag ctgaactacc caaaagtata aatgcctgaa
     6001 caattagttt agatccgagc tagcattaag tcctcagcga gcacgctgct tcatggaatg
     6061 cgtgcgatga gcgacctcat gctatacctg agaaagcaac ctgacctaca ggaaagagtt
     6121 actcaagaat aagaattttc gttttaaaac ctaagagtca ctttaaaatt tgtatacact
     6181 tatttttttt ataacttatt taataataaa aatcataaat cataagaaat tcgcttattt
     6241 agaagtgtca acaacgtatc taccaacgga atg
//
