LOCUS       sequence-116411-        6997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EasyClone system-based yeast integrative vector carrying 
            loxP-flanked KlLEU2 marker, integration into S. cerevisiae XI-2 
            chromosomal location, USER site for cloning, amp resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6997)
  AUTHORS   Stovicek V, Borja GM, Forster J, Borodina I
  TITLE     EasyClone 2.0: expanded toolkit of integrative vectors for stable 
            gene expression in industrial Saccharomyces cerevisiae strains.
  JOURNAL   J Ind Microbiol Biotechnol. 2015 Sep 16.
  PUBMED    26376869
REFERENCE   2  (bases 1 to 6997)
  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: "J Ind 
            Microbiol Biotechnol. 2015 Sep 16."
COMMENT     Sequence Label: pCfB2190
FEATURES             Location/Qualifiers
     source          1..6997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      30..219
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     protein_bind    267..300
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2589..2622
                     /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(2665..2684)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2666..2684)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2694..2710)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2703..2725)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(3376..3409)
                     /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(3879..3895)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3994..4011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4165..4753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4245..4264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4877..5668,5669..5737))
                     /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     5500..5519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5738..5829)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5871..5890
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5896..5915
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6122..6155
                     /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(6803..6968)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6889..6913
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 caacggaatg cgtgcgatcg cgtgcattca tccgctctaa ccgaaaagga aggagttaga
       61 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
      121 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
      181 ctgaaaacct tgcttgagaa ggttttggga cgctcgaaga tcgcgtcagc tgaagcttcg
      241 tacgctgcag gtcgacaacc cttaatataa cttcgtataa tgtatgctat acgaagttat
      301 taggtctaga gatccgcagg ctaaccggaa cctgtattat ttagtttatg ctacgttaaa
      361 taaagacctt tcgttcacat aactgaatgt gtaatggcct tgagatttca agcataccaa
      421 gttggtggag acggggtcgt tacaaaagac tctttaagcc aagatttcct tgacagcctt
      481 ggcgatagca tcgccaacct cagtggtaga gttagaacca ccaaggtcac cggttctgac
      541 acctgcatcc aagacatttc taacagcttc ttcaagagcc ctaccttctt caaccaaatc
      601 caaggataac ttcaacatca tagctgcaga taagatggta gcaattgggt taaccttgtt
      661 tgctggtaaa tctggggcag aaccatgaca tggttcgtac aaaccgaatg ccttgttagt
      721 gtcaggtagg gaagctagag atgcagaagg taataaaccc aaagaacctg gaataacaga
      781 ggcttcatcg gagataatat caccaaacat gttgttggta ataacaacac cgtttagctt
      841 agttggtgat ttaaccaaaa tcatagcagc agagtcgatc aattggtgct gaacagttaa
      901 ttgtgggaac tcagtcttga tggtttcttc aacagtcttt ctccacaatc tggaagaggc
      961 aagcacgtta gccttgtcaa gtgaccagat tggtaatggt gggttttgtt gcaatgccaa
     1021 gaaagcagcc attcttgtaa ttctttgaac ttcaggaaca ctgtatttct cagagtccca
     1081 agcaactccg tcaccttcat cttcttttct ttcaccaaag tagataccac caaccaattc
     1141 tctaacgacg acgaaatcgg taccctttgc atattcaggc ttcaaaggag aaagatctag
     1201 taaagaatca gaagcaaagt tacatggtct taagttggcg tatagaccca attcctttct
     1261 gatcttcaat agaccttgtt ctggtctaac tgcgcccgta ccccattttg gaccaccaac
     1321 agcacctagt aagacagcat cggctttctt agaggcttct agagcttcat ctggtaaagg
     1381 agtgccagtg gcatcgatgg cagcaccccc gatcaagtga tgttggaaat tgaacttaat
     1441 ttctggacgg acttcagcaa tggcattcaa gaccttaata gcttcgtcag taacttcttt
     1501 accgacgtga tcacccggta ggacaacgat attcttagac atggtataat ctgtgtagtg
     1561 tgggatactt tttacttctt caaataggta tcaacaataa aaataaatcg aagcaaatgt
     1621 aggaatgcgt taaagcagat gtacttactt agagtacata gatatattta tataattcaa
     1681 tatataaaag tatatgaagc atctgatgtt gaacctgtca tgactctaaa ctggaatggg
     1741 tagttatgct tctgaaggtt ttccgttaga aaatgagtgc agggatcagg cccggaaccg
     1801 gttttagcct gaaaggaaaa gaaaaaaatg cggtgattac tagcacgtga ctgcgctgaa
     1861 ttggaatcca ccacaagtac agcgaatggc aagagggaag gggaatatat tgtgcctggc
     1921 ggtagtctgc tatatcaaat gttaaacata actaaatgca gtagtgaata gagtaatgta
     1981 tttctacata tgtgggtagg gccaatggga gcccgatgtc tattgtacag atattttcct
     2041 tttatagtta atcaattcaa cttggaaatc attgagtcga tatctacttg gataactgga
     2101 gccgtggcag cgtttgaaat ttgacgagtc tccttgtttt gttcgcgcat tttcagttta
     2161 ttcaaacggg agatggcacc gaatccactg acttctctgt tcaatttgaa tggtaatagg
     2221 aagaaagtac atttcgagaa attttcgccc aacgtcgaaa tctcatggat gatatcctcg
     2281 atacaaatga caccttcatc acctaatttc tcttcaacga tattgttgtc attcaagatt
     2341 atctcttttg gttctttatc ttctggcctt tgccacatta ttctagatct cttttggatt
     2401 aaagaacgga tagatgcaag tgaaggttgt ccgataacaa tatatggtga tacaagtttc
     2461 aacaatgggt acacgtcctt ggtaagtttg atgaaaacac cagtgttcaa ctcaacaagt
     2521 ctcaaaagag ataacacttt gtaagccttt gctgggatct tcacagcgag ctctcgagaa
     2581 cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtgat atcagatcca
     2641 ctagtggcct atgcacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     2701 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccctgcagg actagtgctg
     2761 aggcattaat cacacgacta gcgctttcag atattaaaaa gtttagatgt aggttttagc
     2821 ggtaacagtt atataaatcg tgtttcttct cttgatgaaa caaaaaaatg ctagaaaaac
     2881 tttgtcgttt cttacttttg gtgcgctttg cagttttcgt ggctagactt agaatcattt
     2941 ctcctcagat ttcttgatta aagtttggtg cgaagcccta ctctaacatt ggtgttcttc
     3001 ttttcattca cgcaagttaa gtccaggaag gtgagcaaat gctcatcctt ctgttcatgc
     3061 gtgacggctg aattatcctt atctggcgta cccgtgcagc cgtttccgtg cctcggttcc
     3121 tccgagatat ccttagggac cgccagggac catgattgcg tcaactgttg tcaccgctcc
     3181 agaggatcct ctgtaacctt ttcaaccata aaaatagaaa gggcacaggg ttggggtatt
     3241 tgaatttttt ttttgggttt tttcgtatta cttattactt accttctttt ctatataatt
     3301 ttgtttttcc ctggtagagc ggaatcttcc cacattaaga cctcagcgcg gccgcaaatt
     3361 taaataaaat gaagtgaagt tcctatactt tctagagaat aggaacttct atagtgagtc
     3421 gaataagggc gacacaaaat ttattctaaa tgcataataa atactgataa catcttatag
     3481 tttgtattat attttgtatt atcgttgaca tgtataattt tgatatcaaa aactgatttt
     3541 ccctttatta ttttcgagat ttattttctt aattctcttt aacaaactag aaatattgta
     3601 tatacaaaaa atcataaata atagatgaat agtttaatta taggtgttca tcaatcgaaa
     3661 aagcaacgta tcttatttaa agtgcgttgc ttttttctca tttataaggt taaataattc
     3721 tcatatatca agcaaagtga caggcgccct taaatattct gacaaatgct ctttccctaa
     3781 actcccccca taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg attaacgatt
     3841 actcgttatc agaaccgccc agggggcccg agcttaagac tggccgtcgt tttacaacac
     3901 agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc ttagtttgat
     3961 gcctggcagt tccctactct cgccttccgc ttcctcgctc actgactcgc tgcgctcggt
     4021 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4081 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4501 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4561 tacagagttc ttgaagtggt gggctaacta cggctacact agaagaacag tatttggtat
     4621 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4681 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4741 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4801 cgcgcgcgta actcacgtta agggattttg gtcatgagct tgcgccgtcc cgtcaagtca
     4861 gcgtaatgct ctgcttttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4921 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4981 gcttaccatc tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg
     5041 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5101 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5161 ttaatagttt gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt
     5221 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5281 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5341 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5401 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5461 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5521 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5581 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5641 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5701 agggaataag ggcgacacgg aaatgttgaa tactcatatt cttccttttt caatattatt
     5761 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5821 ataaacaaat aggggtcagt gttacaacca attaaccaat tctgaacatt atcgcgagcc
     5881 catttatacc tgaatatggc tcataacacc ccttgtttgc ctggcggcag tagcgcggtg
     5941 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     6001 gggactcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     6061 gaaagactgg gcctttcgcc cgggctaatt atggggtgtc gcccttattc gactctatag
     6121 tgaagttcct attctctaga aagtatagga acttctgaag tggggattta aatgcggccg
     6181 cgctgagggt ttaatgagga ttttcgatgg agcaggatga ggagaaatag taccacatgt
     6241 atatatccat tacaaaaagg tttatataca attacaatag acccttgttg gggtttctga
     6301 aaaaagaagt agtcgatgcc atcggcaata atacggaatt acgagaaaca caatcccgat
     6361 ccttttttgg gtaattactt caccgattct accgatttat catgccaaaa aaaattcacc
     6421 gtgggttcta gaagtgccct ttgaggattg tagccactct aacccacacg gcctccttac
     6481 tagctgacta aggtgacaaa accgcaagga ctggaaagtc gccactcatc tgaaaattct
     6541 caagtttttc actactgagt ttatgctttc gaattttttt gttcggtaat agcacggcgg
     6601 ttcgattcaa ttccgccgct ccgagcgatg ctccgcaaaa ctcagtaata agctttctga
     6661 tggttcaccc cttttttagc acgcggggtg taactcaaca gaaaaatgtg ccatagaaca
     6721 agactaggca aaagccaagg agcgtttgcc atgaacttcc acaaattaag tcctcagcga
     6781 gctcgcatgg aatgcgtgcg atgagcgacc tcatgctata cctgagaaag caacctgacc
     6841 tacaggaaag agttactcaa gaataagaat tttcgtttta aaacctaaga gtcactttaa
     6901 aatttgtata cacttatttt ttttataact tatttaataa taaaaatcat aaatcataag
     6961 aaattcgctt atttagaagt gtcaacaacg tatctac
//
