LOCUS       sequence-116387-        7022 bp    DNA     circular SYN 14-APR-2025
DEFINITION  EasyClone system-based yeast integrative vector carrying 
            loxP-flanked amdSYM marker, integration into S. cerevisiae XI-5 
            chromosomal location, USER site for cloning, amp resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7022)
  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 7022)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pCfB2399
FEATURES             Location/Qualifiers
     source          1..7022
                     /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    2572..2605
                     /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(2648..2667)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2649..2667)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2677..2693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2686..2708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(3307..3340)
                     /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(3810..3826)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3925..3942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4096..4684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4176..4195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4808..5599,5600..5668))
                     /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     5431..5450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5669..5760)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5802..5821
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5827..5846
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6053..6086
                     /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(6828..6993)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6914..6938
                     /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 gcttgcctcg 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 aaacaaggat ccatgccaca atcttgggaa
      721 gaattggctg ctgacaagag agctagattg gctaagacta tcccagacga atggaaggtt
      781 caaactttgc cagctgaaga ttctgttatc gacttcccaa agaagtctgg tatcttgtct
      841 gaagctgaat tgaagatcac tgaagcttct gctgctgact tggtttctaa gttggctgct
      901 ggtgaattga cttctgttga agttactttg gctttctgta agagagctgc tatcgctcaa
      961 caattgacta actgtgctca cgaattcttc ccagacgctg ctttggctca agctagagaa
     1021 ttggacgaat actacgctaa gcacaaaaga ccagttggtc cattgcacgg tttgccaatc
     1081 tctttgaagg accaattgag agttaagggt tacgaaactt ctatgggtta catctcttgg
     1141 ttgaacaagt acgacgaagg tgactctgtt ttgactacta tgttgagaaa ggctggtgct
     1201 gttttctacg ttaagacttc tgttccacaa actttgatgg tttgtgaaac tgttaacaac
     1261 atcatcggta gaactgttaa cccaagaaac aagaactggt cttgtggtgg ttcttctggt
     1321 ggtgaaggtg ctatcgttgg tatcagaggt ggtgttatcg gtgttggtac tgacatcggt
     1381 ggttctatca gagttccagc tgctttcaac ttcttgtacg gtttaagacc atctcacggt
     1441 agattgccat acgctaagat ggctaactct atggaaggtc aagaaactgt acactctgtt
     1501 gttggtccaa tcactcactc tgttgaagat ttgagattgt tcactaagtc tgttttgggt
     1561 caagaaccat ggaagtacga ctctaaggtt atcccaatgc catggagaca atctgaatct
     1621 gacatcatcg cttctaagat caagaacggt ggtttgaaca tcggttacta caacttcgac
     1681 ggtaacgttt tgccacaccc accaatcttg agaggtgttg aaactactgt tgctgctttg
     1741 gctaaggctg gtcacactgt tactccatgg actccataca agcacgactt cggtcacgac
     1801 ttgatctctc acatctacgc tgctgacggt tctgctgacg ttatgagaga catctctgct
     1861 tctggtgaac cagctatccc aaacatcaag gacttgttga acccaaacat caaggctgtt
     1921 aacatgaacg aattgtggga cactcacttg caaaagtgga actaccaaat ggaatacttg
     1981 gaaaagtgga gagaagctga agaaaaggct ggtaaggaat tggacgctat catcgctcca
     2041 atcactccaa ctgctgctgt tagacacgac caattcagat actacggtta cgcttctgtt
     2101 atcaacttgt tggacttcac ttctgttgtt gttccagtta ctttcgctga caagaacatc
     2161 gacaagaaga acgaatcttt caaggctgtt tctgaattgg acgctttggt tcaagaagaa
     2221 tacgacccag aagcttacca cggtgctcca gttgctgttc aagttatcgg tagaagattg
     2281 tctgaagaaa gaactttggc tatcgctgaa gaagttggta agttgttggg taacgttgtt
     2341 actccataac tcgaggacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     2401 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     2461 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     2521 aatcgtatgt gaatgctggt cgctatactg gagctctcga gaacccttaa tataacttcg
     2581 tataatgtat gctatacgaa gttattaggt gatatcagat ccactagtgg cctatgcacc
     2641 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     2701 tgactgggaa aaccctggcg ttacccctgc aggactagtg ctgaggcatt aatgccttcg
     2761 atttgacaca tctctaagct gaaactgaga atactgttgt aaaacaggta ttggctgctt
     2821 catagtacac ccaattgccc acaacccaag ttaatcataa gctcttgttg tcgatgaatt
     2881 gctcaaaatg tggccattct ttcaattttg cacctctaaa actgctccgt gggttgtgac
     2941 aacaacctta agggtaatgg acttttttag aaagacaaat gagatgattt cacctactgg
     3001 tatatcattt atttatacac gaccgttttg acatttcaga gttgcacgtt ggtatatcct
     3061 aatattctca caagcatcac agtgaagaaa cgcaagaaaa tggaaaaaat tcccaggtgg
     3121 ttgttgtctt ctcttatatc gtcggtgctg tgcatacttg gggccctgtg tgtgccgttg
     3181 ttatcggttg ccttcgatag caagcgcaat agccaatcta agttggtcaa ctatggtctt
     3241 tctctaagtg ccggatctat gatcattaag acctcagcgc ggccgcaaat ttaaataaaa
     3301 tgaagtgaag ttcctatact ttctagagaa taggaacttc tatagtgagt cgaataaggg
     3361 cgacacaaaa tttattctaa atgcataata aatactgata acatcttata gtttgtatta
     3421 tattttgtat tatcgttgac atgtataatt ttgatatcaa aaactgattt tccctttatt
     3481 attttcgaga tttattttct taattctctt taacaaacta gaaatattgt atatacaaaa
     3541 aatcataaat aatagatgaa tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt
     3601 atcttattta aagtgcgttg cttttttctc atttataagg ttaaataatt ctcatatatc
     3661 aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc tctttcccta aactcccccc
     3721 ataaaaaaac ccgccgaagc gggtttttac gttatttgcg gattaacgat tactcgttat
     3781 cagaaccgcc cagggggccc gagcttaaga ctggccgtcg ttttacaaca cagaaagagt
     3841 ttgtagaaac gcaaaaaggc catccgtcag gggccttctg cttagtttga tgcctggcag
     3901 ttccctactc tcgccttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3961 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4501 cttgaagtgg tgggctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgcgcgt
     4741 aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc
     4801 tctgctttta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4861 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4921 ctggccccag cgctgcgatg ataccgcgag aaccacgctc accggctccg gatttatcag
     4981 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5041 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5101 tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5161 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5221 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5281 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5341 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5401 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5461 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5521 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5581 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5641 gggcgacacg gaaatgttga atactcatat tcttcctttt tcaatattat tgaagcattt
     5701 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5761 taggggtcag tgttacaacc aattaaccaa ttctgaacat tatcgcgagc ccatttatac
     5821 ctgaatatgg ctcataacac cccttgtttg cctggcggca gtagcgcggt ggtcccacct
     5881 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggactccc
     5941 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     6001 ggcctttcgc ccgggctaat tatggggtgt cgcccttatt cgactctata gtgaagttcc
     6061 tattctctag aaagtatagg aacttctgaa gtggggattt aaatgcggcc gcgctgaggg
     6121 tttaatgcgg agaagtcgtt gatagcattt ccgaaggctt ttccacattg actgcagtat
     6181 ctaaaagagc tccagcagcg gtcttagtag ttcctgtgct tgccgctttg cagtatacag
     6241 ggacattggg aactacggag gaaaaatcgg aatgctctga gctcttctca aaaccagacg
     6301 aactcgtctt atttctcagt aaattaccca acatataaac tctccctctt caccttctct
     6361 acaaggccac acacaagaca gttttaacag gcaaatggta ctggaggctc tcaccaatgg
     6421 ggtttaagac gctccaaaca cacgctctcc tttttttctt tatatatcgg tagcaacgaa
     6481 agctagtcgc agaacgttac acactaaagc agatcaatca atcaatcctg agaacctgct
     6541 tcgttcaaac catccactag aagaaacacg cacgtgtacc caaaaataga gacaaacaaa
     6601 cagacaaaaa taacaatgat tcctaccgac taaagtaaaa caacactcac gtaataaaaa
     6661 atcctagata caactcactt gtattcctct cttctttgac catcactaaa gcttctctct
     6721 tactatattc ctcaaataaa aaaaagttgc aatgattact taccaatgtg ccataaactc
     6781 cgtgcaccaa ttaagtcctc agcgagctcg catggaatgc gtgcgatgag cgacctcatg
     6841 ctatacctga gaaagcaacc tgacctacag gaaagagtta ctcaagaata agaattttcg
     6901 ttttaaaacc taagagtcac tttaaaattt gtatacactt atttttttta taacttattt
     6961 aataataaaa atcataaatc ataagaaatt cgcttattta gaagtgtcaa caacgtatct
     7021 ac
//
