LOCUS       40924_10431        6222 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning and expression vector pCEV-G3-Ph, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6222)
  AUTHORS   Vickers CE, Bydder SF, Zhou Y, Nielsen LK.
  TITLE     Dual gene expression cassette vectors with antibiotic selection 
            markers for engineering in Saccharomyces cerevisiae
  JOURNAL   Microb. Cell Fact. 12 (1), 96 (2013)
  PUBMED    24161108
REFERENCE   2  (bases 1 to 6222)
  AUTHORS   Bydder SF, Vickers CE.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUL-2013) AIBN, The University of Queensland, Crn 
            College and Cooper Roads (Bldg 75), St Lucia, QLD 4072, Australia
REFERENCE   3  (bases 1 to 6222)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6222)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2013"; volume: "12"; issue: "1"; pages: "96"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JUL-2013) AIBN, The University of Queensland, Crn College and 
            Cooper Roads (Bldg 75), St Lucia, QLD 4072, Australia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2..167)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(315..338)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        356..1008
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        1017..1424
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1444..1462
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1480..1509
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      1539..1728
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(1971..2559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2733..3590)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3591..3695)
                     /label=AmpR promoter
     rep_origin      complement(3722..5064)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5084..5117
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5172..5515
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5516..5902
                     /codon_start=1
                     /gene="ble"
                     /product="bleomycin resistance protein"
                     /label=ble
                     /protein_id="AGZ03675.1"
                     /translation="MGMTDQATPNLPSRDFDPTAAFYERLGFGIVFRDAGWMILQRGDL
                     MLEFFAHPGLDPLASWFSCCLRLDDLAEFYRQCKSVGIQETSSGYPRIHAPELQEWGGT
                     MAALVDPDGTLLRLIQNELLAGIS"
     gene            5516..5902
                     /gene="ble"
                     /label=ble
     terminator      5908..6105
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_recomb     6168..6201
                     /label=loxP recognition sequence for Cre recombinase
                     /note="loxP recognition sequence for Cre recombinase"
     protein_bind    complement(6168..6201)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
ORIGIN
        1 ggagcgacct catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag
       61 aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt
      121 tttataactt atttaataat aaaaatcata aatcataaga aattcgctta tttagaagtg
      181 tcaacaacgt atctaccaac gatttgaccc ttttccatct tttcgtaaat ttctggcaag
      241 gtagacaagc cgacaacctt gattggagac ttgaccaaac ctctggcgaa gaattgttaa
      301 ttaagagctc agatcttatc gtcgtcatcc ttgtaatcca tcgatactag tgaattttca
      361 aaaattctta cttttttttt ggatggacgc aaagaagttt aataatcata ttacatggca
      421 ttaccaccat atacatatcc atatctaatc ttacttatat gttgtggaaa tgtaaagagc
      481 cccattatct tagcctaaaa aaaccttctc tttggaactt tcagtaatac gcttaactgc
      541 tcattgctat attgaagtac ggattagaag ccgccgagcg ggcgacagcc ctccgacgga
      601 agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct
      661 cgcgccgcac tgctccgaac aataaagatt ctacaatact agcttttatg gttatgaaga
      721 ggaaaaattg gcagtaacct ggccccacaa accttcaaat taacgaatca aattaacaac
      781 cataggatga taatgcgatt agttttttag ccttatttct ggggtaatta atcagcgaag
      841 cgatgatttt tgatctatta acagatatat aaatggaaaa gctgcataac cactttaact
      901 aatactttca acattttcag tttgtattac ttcttattca aatgtcataa aagtatcaac
      961 aaaaaattgt taatatacct ctatacttta acgtcaagga gaaaaaacgg ccggccgcac
     1021 acaccatagc ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc
     1081 gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttc ccctctttct
     1141 tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc
     1201 tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg
     1261 aaaatttttt tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat
     1321 aaacggtctt caatttctca agtttcagtt tcatttttct tgttctatta caactttttt
     1381 tacttcttgc tcattagaaa gaaagcatag caatctaatc taagttttaa ttacaaggat
     1441 ccgtaatacg actcactata gggcccgggc gtcgacatgg aacagaagtt gatttccgaa
     1501 gaagacctcg agtaagcttg gtaccgcggc tagctaagat ccgctctaac cgaaaaggaa
     1561 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     1621 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     1681 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaagat ccagctgcat
     1741 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1801 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2401 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2521 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2641 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2701 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2761 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2821 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2881 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2941 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3001 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3061 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3121 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3181 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3241 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3301 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3361 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3421 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3481 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3541 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3721 gaacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     3781 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     3841 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     3901 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     3961 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     4021 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     4081 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     4141 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     4201 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     4261 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     4321 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     4381 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     4441 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     4501 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     4561 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     4621 ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg
     4681 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tcaaagcgtt
     4741 tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca
     4801 cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt
     4861 gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg aaaggtagtc
     4921 tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc cttcagcact
     4981 accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat ccttcaatgc
     5041 tatcatttcc tttgatattg gatcatggta gacaaccctt aatataactt cgtataatgt
     5101 atgctatacg aagttattag gtctagagat ctgtttagct tgcctcgtcc ccgccgggtc
     5161 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
     5221 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
     5281 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
     5341 agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
     5401 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
     5461 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg
     5521 tatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgatccca ccgccgcctt
     5581 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5641 cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg cgagttggtt
     5701 cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca aatccgtcgg
     5761 catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc aggagtgggg
     5821 aggcacgatg gccgctttgg tcgacccgga cgggacgctc ctgcgcctga tacagaacga
     5881 attgcttgca ggcatctcat gatcagtact gacaataaaa agattcttgt tttcaagaac
     5941 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     6001 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     6061 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
     6121 taacgccgcc atccagtgtc gaaaacgagc tctcgagaac ccttaatata acttcgtata
     6181 atgtatgcta tacgaagtta ttaggtgata tcagatccac ta
//
