LOCUS       sequence-124525-        6113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EasyClone system-based yeast integrative vector carrying 
            loxP-flanked KlURA3 marker, integration into S. cerevisiae XI-1 
            chromosomal location, USER site for cloning, amp resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6113)
  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 6113)
  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: pCfB2374
FEATURES             Location/Qualifiers
     source          1..6113
                     /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    1753..1786
                     /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(1829..1848)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1830..1848)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1858..1874)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1867..1889)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(2600..2633)
                     /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(3103..3119)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3218..3235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3389..3977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3469..3488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4101..4892,4893..4961))
                     /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     4724..4743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4962..5053)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5095..5114
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5120..5139
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5346..5379
                     /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(5919..6084)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6005..6029
                     /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 gatcccaata caacagatca cgtgatcttt ttgtaagatg aagttgaagt
      361 agtgttgcac cgtgccaatg caggtggcta ttagattaaa tatgtgattt gttctattaa
      421 gtttcctgta taattaatgg ggagcgctga ttctcttttg gtacgcttcc catccagcat
      481 ttctgtatct ttcaccttca accttaggat ctctaccctt ggcgaaaagt cctctgccaa
      541 caatgatgat atctgatcca ccacttacaa cttcgtcgac ggttctgtac tgctgaccca
      601 atgcatcgcc tttgtcgtct aaacctacac ctggggtcat gattagccaa tcaaaccctt
      661 cttctcttcc tcccatatcg ttctgagcaa tgaacccaat aacgaaatct ttatcactct
      721 ttgcaatatc aacggtaccc ttagtatatt caccgtgtgc tagagaaccc ttggaagata
      781 attcagcaag catcaataat ccccttggtt ctttggtgac ctcttgcgca ccttgtttca
      841 agccagcaac aataccagca ccagtaaccc cgtgggcgtt ggtgatatca gaccattctg
      901 cgatacggta aacgcccgat gtatattgta atttgactgt gttaccgata tcggcgaatt
      961 ttctgtcctc aaatatcaag aacttgtatt tctctgccaa tgctttcaat ggaacgacag
     1021 taccctcata actgaaatca tccaagatat caacgtgtgt tttcaaaagg caaatgtatg
     1081 gacccaacgt ttcaacaagt ttcaatagct catcagtcga acgaacgtca agagaagcac
     1141 acaaattggt tttcttttca tccattaaac gtaaaagttt cgatgcaacc ggacttgcat
     1201 gagtctcagc tctactggta tatgattttg tggacatggt gcaactaatt gacgggagtg
     1261 tattgacgct ggcgtactgg ctttcacaaa atggcccaat cacaaccaca tcttagatag
     1321 ttgaaatgac tttagataac atcaattgag atgagcttaa tcatgtcaaa gctaaaagtg
     1381 tcaccatgaa cgacaattct taagcaaatc acgtgatata gatccacgaa taaccaccat
     1441 ttgatgctcg aggcaagtaa tgtgtgtaaa aaaatgcgtt accaccatcc aatgcagacc
     1501 gatcttctac ccagaatcac atatatttat gtaccgagta ccttttttct atcttccaat
     1561 tgcttctccc atatgattgt ctccgtaagc tcgaaatttc taagttggat tttaatcttc
     1621 acgcaggatg acagttcgat gagcttctga ggagtgttta gaacataatc agtttatcca
     1681 tggtctatct cttcttgtcg ctttttctcc tcgatagaac ctaaataaaa cgagctctcg
     1741 agaaccctta atataacttc gtataatgta tgctatacga agttattagg tgatatcaga
     1801 tccactagtg gcctatgcac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     1861 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccctg caggactagt
     1921 gctgaggcat taatcttcca cggaatacca agcccattgc aatgcgatgt tagtttagtg
     1981 gagtttcttg gcattggcaa atctctgcta aatgctgcgt acagacggaa actcacaccg
     2041 ccgcgaaaac tggtcagtgg caaaaaaaaa aaaaattaaa aaaataaaaa ataactatta
     2101 cgtatgatac tgtttctggt agttgatatg aggttggtgt tgtatattgt acgttttagg
     2161 aacagggaag tgaatattat ttactctgct gcacattctg gctaggtcga agccggaacc
     2221 tgagatgacg ccgcgctaga actatggacc aagctgttga caatgtttag atggtgatgc
     2281 actaccctgt gcggggagtg gccacgaacg cgagcggaag gtgcgggtgt tacgggaatt
     2341 gcgggaggtt cttcgctaag cgtgagggtt gctagttggg gcggcggggt ttccctaagt
     2401 gtaaataggg cctcgccgct ggcacatgag tgcgccggag gaggcggcgg aggcgacgac
     2461 gctaaaaccg tggccgttgg ggaaggatgg gcggctatat ctaccattga cctgatgggg
     2521 actcggctct taaggaatgg gtttgaggtg ggtgtggatt aagacctcag cgcggccgca
     2581 aatttaaata aaatgaagtg aagttcctat actttctaga gaataggaac ttctatagtg
     2641 agtcgaataa gggcgacaca aaatttattc taaatgcata ataaatactg ataacatctt
     2701 atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga
     2761 ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat
     2821 tgtatataca aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc
     2881 gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata
     2941 attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc
     3001 ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac
     3061 gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca
     3121 acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt
     3181 tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct
     3241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3781 gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga acagtatttg
     3841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4021 acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     4081 gtcagcgtaa tgctctgctt ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4201 gagggcttac catctggccc cagcgctgcg atgataccgc gagaaccacg ctcaccggct
     4261 ccggatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4321 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4381 ccagttaata gtttgcgcaa cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg
     4441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tattcttcct ttttcaatat
     4981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5041 aaaaataaac aaataggggt cagtgttaca accaattaac caattctgaa cattatcgcg
     5101 agcccattta tacctgaata tggctcataa caccccttgt ttgcctggcg gcagtagcgc
     5161 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     5221 tgtggggact ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     5281 agtcgaaaga ctgggccttt cgcccgggct aattatgggg tgtcgccctt attcgactct
     5341 atagtgaagt tcctattctc tagaaagtat aggaacttct gaagtgggga tttaaatgcg
     5401 gccgcgctga gggtttaatg gaatagtgac gttgtgatgc ggtgagttcg gcggttaggg
     5461 gaatggtata tgataaaaaa cggaaacgtg cttctttaat ttaattgttt aatattgttg
     5521 cagatatata aaaaggggga aagaaccgaa gatgtaatta tttttttatc gcctcaacct
     5581 aaagcaagca ataaggtata aagatcagga cgtctcgagc gctgatatct aaatttgaag
     5641 ccacgcaagt aactacgtag gtcagagggg acaaggaata acacttgaca tttttctttt
     5701 ttcttttttt ttcttttttt tttttttgtt aatcttggct tctgtaccgt agcctcctgt
     5761 atacggtttt agatgcagta tgtttatcat cgccgagaaa tttggggggt gcaaaggaat
     5821 aaaatccgtg ctgaaatccg tgcaccgcat caaactttct cggaggattc tttgcagccg
     5881 attaagtcct cagcgagctc gcatggaatg cgtgcgatga gcgacctcat gctatacctg
     5941 agaaagcaac ctgacctaca ggaaagagtt actcaagaat aagaattttc gttttaaaac
     6001 ctaagagtca ctttaaaatt tgtatacact tatttttttt ataacttatt taataataaa
     6061 aatcataaat cataagaaat tcgcttattt agaagtgtca acaacgtatc tac
//
