LOCUS       sequence-148849-        5444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCfB2797 is a vector for multiple integrations at sites sharing 
            homology with Ty2Cons. 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 5444)
  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 5444)
  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: pCfB2797
FEATURES             Location/Qualifiers
     source          1..5444
                     /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(2180..2213)
                     /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(2683..2699)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2798..2815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2969..3557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3049..3068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3681..4472,4473..4541))
                     /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     4304..4323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4542..4633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4675..4694
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4700..4719
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    4926..4959
                     /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(5240..5405)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5326..5350
                     /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 tacccctgca ggacagacgt cattagtgct gaggcattaa
     1981 ttgatcaata taaaatgatg ataataatat ttatagaatt gtgtagaatt gcagattccc
     2041 ttttatggat tcctaaatcc tgaggagaac ttctagtata ttctacatac ctaatattat
     2101 tgccttatta aaaatggaat cccaacaatt acatcaaaat ccacattctc agatctgcgg
     2161 ccgcactcag acctgaagtg aagttcctat actttctaga gaataggaac ttctatagtg
     2221 agtcgaataa gggcgacaca aaatttattc taaatgcata ataaatactg ataacatctt
     2281 atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga
     2341 ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat
     2401 tgtatataca aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc
     2461 gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata
     2521 attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc
     2581 ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac
     2641 gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca
     2701 acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt
     2761 tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct
     2821 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2881 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2941 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3001 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3061 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3121 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3181 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3241 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3301 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3361 gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga acagtatttg
     3421 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3481 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3541 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3601 acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     3661 gtcagcgtaa tgctctgctt ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3781 gagggcttac catctggccc cagcgctgcg atgataccgc gagaaccacg ctcaccggct
     3841 ccggatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3961 ccagttaata gtttgcgcaa cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg
     4021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tattcttcct ttttcaatat
     4561 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4621 aaaaataaac aaataggggt cagtgttaca accaattaac caattctgaa cattatcgcg
     4681 agcccattta tacctgaata tggctcataa caccccttgt ttgcctggcg gcagtagcgc
     4741 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     4801 tgtggggact ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     4861 agtcgaaaga ctgggccttt cgcccgggct aattatgggg tgtcgccctt attcgactct
     4921 atagtgaagt tcctattctc tagaaagtat aggaacttct gaagtgggga tttaaagtcg
     4981 gtgtccgcgc tgagggttta atggcgcgcc gcggccgccc gcggtgttgg aataaaaatc
     5041 aactatcatc tactaactag tatttacgtt actagtatat tatcatatac ggtgttagaa
     5101 gatgacgcaa atgatgagaa atagtcatct aaattagtgg aagctgaaac gcaaggattg
     5161 ataatgtaat aggatcaatg aatattaacg ctagcattaa gtcctcagcg agcacgctgc
     5221 ttcatggaat gcgtgcgatg agcgacctca tgctatacct gagaaagcaa cctgacctac
     5281 aggaaagagt tactcaagaa taagaatttt cgttttaaaa cctaagagtc actttaaaat
     5341 ttgtatacac ttattttttt tataacttat ttaataataa aaatcataaa tcataagaaa
     5401 ttcgcttatt tagaagtgtc aacaacgtat ctaccaacgg aatg
//
