LOCUS       sequence-116385-        6670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EasyClone system-based yeast integrative vector carrying 
            loxP-flanked dsdA marker, integration into S. cerevisiae XII-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 6670)
  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 6670)
  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: pCfB2400
FEATURES             Location/Qualifiers
     source          1..6670
                     /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)."
     promoter        355..698
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2033..2230
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    2293..2326
                     /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(2367..2386)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2368..2386)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2396..2412)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2405..2427)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(3104..3137)
                     /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(3607..3623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3722..3739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3893..4481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3973..3992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4605..5396,5397..5465))
                     /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     5228..5247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5466..5557)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5599..5618
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5624..5643
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5850..5883
                     /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(6476..6641)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6562..6586
                     /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 gatctgttta gcttgccttg tccccgccgg gtcacccggc cagcgacatg
      361 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
      421 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
      481 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
      541 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
      601 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
      661 ggatacagtt ctcacatcac atccgaacat aaacaaccat ggaaaacgct aaaatgaact
      721 cgctcatcgc ccagtatccg ttggtaaagg atctggttgc tcttaaagaa accacctggt
      781 ttaatcctgg cacgacctca ttggctgaag gtttacctta tgttggcctg accgaacagg
      841 atgttcagga cgcccatgcg cgcttatccc gttttgcacc ctatctggca aaagcatttc
      901 ctgaaactgc tgccactggg gggattattg aatcagaact ggttgccatt ccagctatgc
      961 aaaaacggct ggaaaaagaa tatcagcaac cgatcagcgg gcaactgtta ctgaaaaaag
     1021 atagccattt gcccatttcc ggctccataa aagcacgcgg cgggatttat gaagtcctgg
     1081 cacacgcaga aaaactggct ctggaagcgg ggttgctgac gcttgatgat gactacagca
     1141 aactgctttc tccggagttt aaacagttct ttagccaata cagcattgct gtgggctcaa
     1201 ccggaaatct ggggttatca atcggcatta tgagcgcccg cattggcttt aaggtgacag
     1261 ttcatatgtc tgctgatgcc cgggcatgga aaaaagcgaa actgcgcagc catggcgtta
     1321 cggtcgtgga atatgagcaa gattatggtg ttgccgtcga ggaaggacgt aaagcagcgc
     1381 agtctgaccc gaactgtttc tttattgatg acgaaaattc ccgcacgttg ttccttgggt
     1441 attccgtcgc tggccagcgt cttaaagcgc aatttgccca gcaaggccgt atcgtcgatg
     1501 ctgataaccc tctgtttgtc tatctgccgt gtggtgttgg cggtggtcct ggtggcgtcg
     1561 ctttcgggct taaactggcg tttggcgatc atgttcactg cttttttgcc gaaccaacgc
     1621 actccccttg tatgttgtta ggcgtccata caggattaca cgatcagatt tctgttcagg
     1681 atattggtat cgacaacctt accgcagcgg atggccttgc agttggtcgc gcatcaggct
     1741 ttgtcgggcg ggcaatggag cgtctgctgg atggcttcta tacccttagc gatcaaacca
     1801 tgtatgacat gcttggctgg ctggcgcagg aagaaggtat tcgtcttgaa ccttcggcac
     1861 tggcgggtat ggccggacct cagcgcgtgt gtgcatcagt aagttaccaa cagatgcacg
     1921 gtttcagcgc agaacaactg cgtaatacca ctcatctggt gtgggcgacg ggaggtggaa
     1981 tggtgccgga agaagagatg aatcaatatc tggcaaaagg ccgttaatca gtactgacaa
     2041 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     2101 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     2161 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     2221 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctctcg
     2281 agaaccctta atataacttc gtataatgta tgctatacga agttattagg tgatatcaga
     2341 tccactagtg gcctataccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     2401 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccctgca ggactagtgc
     2461 tgaggcatta atcttcccgt gaatcaactg cacatacctt aataactagt tcagtttagt
     2521 gctctgtctg agtgactgta ataataaatt ttacaagcca cttctcatga catatattgg
     2581 taagtaactt catcaatact aattagtctt tgccggttac ccatctggcc cctgacttgc
     2641 gatgcttagg aagttccata ctcgcggctc ttcccaacag tagcacatcc gtgaaacttc
     2701 tggcgctatt cattatgcag taccaggaca agaagttaaa aaaaaagctc tgttacaagt
     2761 tcaatggtgg tgcaaggatt gaagttatta tccaggaggc acggatgcaa aagatagagg
     2821 acaaattaat ttccttaaaa ttcaaaatgc tcattattgt cacgctgtat gagcatttgg
     2881 tgaagatttc actgggaaat gttgcaataa tttgataatc gttcgtattg gatgaaactg
     2941 taacatcatc tgtttattaa gtatccgtgt tattagtata tcatcacata cggtgtaaga
     3001 agataacata aagattgaga aacagtcatc aaatataatg gaagctgaaa tgcgaggatt
     3061 gattaagacc tcagcgcggc cgcaaattta aataaaatga agtgaagttc ctatactttc
     3121 tagagaatag gaacttctat agtgagtcga ataagggcga cacaaaattt attctaaatg
     3181 cataataaat actgataaca tcttatagtt tgtattatat tttgtattat cgttgacatg
     3241 tataattttg atatcaaaaa ctgattttcc ctttattatt ttcgagattt attttcttaa
     3301 ttctctttaa caaactagaa atattgtata tacaaaaaat cataaataat agatgaatag
     3361 tttaattata ggtgttcatc aatcgaaaaa gcaacgtatc ttatttaaag tgcgttgctt
     3421 ttttctcatt tataaggtta aataattctc atatatcaag caaagtgaca ggcgccctta
     3481 aatattctga caaatgctct ttccctaaac tccccccata aaaaaacccg ccgaagcggg
     3541 tttttacgtt atttgcggat taacgattac tcgttatcag aaccgcccag ggggcccgag
     3601 cttaagactg gccgtcgttt tacaacacag aaagagtttg tagaaacgca aaaaggccat
     3661 ccgtcagggg ccttctgctt agtttgatgc ctggcagttc cctactctcg ccttccgctt
     3721 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3781 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3841 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3901 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3961 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4021 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4081 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4141 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4201 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4261 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg gctaactacg
     4321 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4381 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4441 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4501 ctacggggtc tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag ggattttggt
     4561 catgagcttg cgccgtcccg tcaagtcagc gtaatgctct gcttttacca atgcttaatc
     4621 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4681 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagcgc tgcgatgata
     4741 ccgcgagaac cacgctcacc ggctccggat ttatcagcaa taaaccagcc agccggaagg
     4801 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4861 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct
     4921 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4981 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5041 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5101 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5161 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5221 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5281 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5341 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5401 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5461 ctcatattct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5521 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt tacaaccaat
     5581 taaccaattc tgaacattat cgcgagccca tttatacctg aatatggctc ataacacccc
     5641 ttgtttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
     5701 gaaacgccgt agcgccgatg gtagtgtggg gactccccat gcgagagtag ggaactgcca
     5761 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgcccg ggctaattat
     5821 ggggtgtcgc ccttattcga ctctatagtg aagttcctat tctctagaaa gtataggaac
     5881 ttctgaagtg gggatttaaa tgcggccgcg ctgagggttt aatgttgagc tctgtccttc
     5941 atggactttt tggaccactt cttcttggac ttcttaccac cagcaagagc ggcagcggct
     6001 ttggcagctt tagacaattg ttgctttgga ggcatgttat attatgttct gagacgtaag
     6061 aaagggtgaa aattgatgtt agtgtcaaaa attatattac aaaatacgca gagatattct
     6121 agttcctttg atgaatgaat ctttcagaaa aaaaagtcaa agcaaaagca aaatggcctg
     6181 cagactaaac tgtatggtgg tcttggaatg ataaagatct gtttaataga tttagtagat
     6241 acaatagcac atctcattac ccagttatga ttgacgtcat tctgagttac aatgatctta
     6301 gcgacctgtg ctcattattt gctccactaa ttctaatttt cctcgccttt catatttcgt
     6361 atctttattc tatatcctaa aatttttttg gcaaatccca gatttggctt tgattttggc
     6421 atcggttcgg ttctttcatt aagtcctcag cgagctcgca tggaatgcgt gcgatgagcg
     6481 acctcatgct atacctgaga aagcaacctg acctacagga aagagttact caagaataag
     6541 aattttcgtt ttaaaaccta agagtcactt taaaatttgt atacacttat tttttttata
     6601 acttatttaa taataaaaat cataaatcat aagaaattcg cttatttaga agtgtcaaca
     6661 acgtatctac
//
