LOCUS       sequence-148863-        6272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2992 is a vector for multiple integrations at sites sharing 
            homology with Ty1Cons1. 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 6272)
  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 6272)
  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: pCfB2992
FEATURES             Location/Qualifiers
     source          1..6272
                     /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(3003..3036)
                     /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(3506..3522)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3621..3638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3792..4380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3872..3891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4504..5295,5296..5364))
                     /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     5127..5146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5365..5456)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5498..5517
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5523..5542
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5749..5782
                     /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(6068..6233)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6154..6178
                     /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 tcaaaaacgg aatgaggaat
     2821 aatcgtaata ttagtatgta gaaatataga ttccattttg aggattccta tatcctcgag
     2881 gagaacttct agtgtatatt ctgtatacct aatattatag cctttatcaa caatggaatc
     2941 ccaacaatta tctaattacc cacaaatttc tcaagatctg cggccgcact cagacctgaa
     3001 gtgaagttcc tatactttct agagaatagg aacttctata gtgagtcgaa taagggcgac
     3061 acaaaattta ttctaaatgc ataataaata ctgataacat cttatagttt gtattatatt
     3121 ttgtattatc gttgacatgt ataattttga tatcaaaaac tgattttccc tttattattt
     3181 tcgagattta ttttcttaat tctctttaac aaactagaaa tattgtatat acaaaaaatc
     3241 ataaataata gatgaatagt ttaattatag gtgttcatca atcgaaaaag caacgtatct
     3301 tatttaaagt gcgttgcttt tttctcattt ataaggttaa ataattctca tatatcaagc
     3361 aaagtgacag gcgcccttaa atattctgac aaatgctctt tccctaaact ccccccataa
     3421 aaaaacccgc cgaagcgggt ttttacgtta tttgcggatt aacgattact cgttatcaga
     3481 accgcccagg gggcccgagc ttaagactgg ccgtcgtttt acaacacaga aagagtttgt
     3541 agaaacgcaa aaaggccatc cgtcaggggc cttctgctta gtttgatgcc tggcagttcc
     3601 ctactctcgc cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3661 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3721 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3781 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3841 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3901 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3961 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4021 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4081 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4141 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4201 aagtggtggg ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4261 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4321 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4381 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgacgc gcgcgtaact
     4441 cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg
     4501 cttttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4561 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4621 ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt tatcagcaat
     4681 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4741 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4801 caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4861 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4921 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4981 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5041 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5101 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5161 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5221 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5281 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5341 gacacggaaa tgttgaatac tcatattctt cctttttcaa tattattgaa gcatttatca
     5401 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5461 ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat ttatacctga
     5521 atatggctca taacacccct tgtttgcctg gcggcagtag cgcggtggtc ccacctgacc
     5581 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg actccccatg
     5641 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     5701 tttcgcccgg gctaattatg gggtgtcgcc cttattcgac tctatagtga agttcctatt
     5761 ctctagaaag tataggaact tctgaagtgg ggatttaaag tcggtgtccg cgctgagggt
     5821 ttaatggcgc gccgcggccg cccgcggtgt tggaataaaa atccactatc gtctatcaac
     5881 taatagttat attatcaata tattatcata tacggtgtta agatgatgac ataagttatg
     5941 agaagctgtc atcgaagtta gaggaagctg aagtgcaagg attgataatg taataggatc
     6001 aatgaatata aacatatgct agcattaagt cctcagcgag cacgctgctt catggaatgc
     6061 gtgcgatgag cgacctcatg ctatacctga gaaagcaacc tgacctacag gaaagagtta
     6121 ctcaagaata agaattttcg ttttaaaacc taagagtcac tttaaaattt gtatacactt
     6181 atttttttta taacttattt aataataaaa atcataaatc ataagaaatt cgcttattta
     6241 gaagtgtcaa caacgtatct accaacggaa tg
//
