LOCUS       sequence-124518-        6049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EasyClone system-based yeast integrative vector carrying 
            loxP-flanked KlURA3 marker, integration into S. cerevisiae X-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 6049)
  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 6049)
  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: pCfB2188
FEATURES             Location/Qualifiers
     source          1..6049
                     /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    1752..1785
                     /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(1828..1847)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1829..1847)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1857..1873)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1866..1888)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(2466..2499)
                     /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(2969..2985)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3084..3101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3255..3843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3335..3354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3967..4758,4759..4827))
                     /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     4590..4609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4828..4919)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4961..4980
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4986..5005
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5212..5245
                     /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(5855..6020)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5941..5965
                     /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 atttatccat
     1681 ggtctatctc ttcttgtcgc tttttctcct cgatagaacc taaataaaac gagctctcga
     1741 gaacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt gatatcagat
     1801 ccactagtgg cctatgcacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     1861 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccctgc aggactagtg
     1921 ctgaggcatt aatcctgcat aatcggcctc acagagggat cccgttaccc atctatgctg
     1981 aagatttatc atactattcc tccgctcgtt tcttttttca gtgaggtgtg tcgtgaaaga
     2041 aaacccacaa ttaaactttc aacaaccggg cgactaggaa gagagtagtg ggcgcggatg
     2101 acgaaggcta aggtcacttc ttcgtttcct ttattggggt ttccgtgtag ccttcccctg
     2161 aatagtgtgg gacgttttat gagaagccgt aagaaatagg caaattgagt tatgacaagt
     2221 agacatgatg ccgcagcctt gcctgacttt acgtctcctt catgaataag tttttctatc
     2281 gagttctttt ccttttttcg ccttaattag ctcaattaag cctgtcctca ctacttttct
     2341 ttttcttatc ggctttgtgc cacacctaac cttcgaatgc tgttttattc cgttcttaca
     2401 tgggatggta atgccttggc gagattaaga cctcagcgcg gccgcaaatt taaataaaat
     2461 gaagtgaagt tcctatactt tctagagaat aggaacttct atagtgagtc gaataagggc
     2521 gacacaaaat ttattctaaa tgcataataa atactgataa catcttatag tttgtattat
     2581 attttgtatt atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta
     2641 ttttcgagat ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa
     2701 atcataaata atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta
     2761 tcttatttaa agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca
     2821 agcaaagtga caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca
     2881 taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc
     2941 agaaccgccc agggggcccg agcttaagac tggccgtcgt tttacaacac agaaagagtt
     3001 tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc ttagtttgat gcctggcagt
     3061 tccctactct cgccttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3121 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3181 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3241 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3301 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3361 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3421 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3481 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3541 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3601 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3661 ttgaagtggt gggctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3721 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3781 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3841 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgcgcgta
     3901 actcacgtta agggattttg gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct
     3961 ctgcttttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4081 tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg atttatcagc
     4141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4261 gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4801 ggcgacacgg aaatgttgaa tactcatatt cttccttttt caatattatt gaagcattta
     4861 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4921 aggggtcagt gttacaacca attaaccaat tctgaacatt atcgcgagcc catttatacc
     4981 tgaatatggc tcataacacc ccttgtttgc ctggcggcag tagcgcggtg gtcccacctg
     5041 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggactcccc
     5101 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     5161 gcctttcgcc cgggctaatt atggggtgtc gcccttattc gactctatag tgaagttcct
     5221 attctctaga aagtatagga acttctgaag tggggattta aatgcggccg cgctgagggt
     5281 ttaatcgtct atgaggagac tgttagttgg atatcagtaa tgagacgaaa aagctcgaaa
     5341 tgaatggata tattcttttt gctactggca actgttgaat atttaatgtt aaaacaaact
     5401 aactgaggta tattcgtatc tgtatgtaca catatactat atacaggaaa agataagcaa
     5461 gagagaggat atcaactacg agagcgatcg attatatatc aaaagctgtc cgctttgcca
     5521 cccataatcg gcgcttagtt tcggagttca atcataattc taccacctta cactcaactt
     5581 actctttaac tcctatagta taatatcgcc actgacccca tattaaaaaa tttttttgct
     5641 cgatcttcta tcctctttag gttaattgtc gctgttattg tctagatttt ttctcggaga
     5701 tggcgcatct atttgccgtc aaaagatcct ctcataccat attaagtaaa ttgcctccat
     5761 ttctttttcc tcgggcagag aaactcgcag gcaacttgct ctcgaagtgg tcacgtatta
     5821 agtcctcagc gagctcgcat ggaatgcgtg cgatgagcga cctcatgcta tacctgagaa
     5881 agcaacctga cctacaggaa agagttactc aagaataaga attttcgttt taaaacctaa
     5941 gagtcacttt aaaatttgta tacacttatt ttttttataa cttatttaat aataaaaatc
     6001 ataaatcata agaaattcgc ttatttagaa gtgtcaacaa cgtatctac
//
