LOCUS       40924_45748        5901 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUMRI-13, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5901)
  AUTHORS   Xie W, Lv X, Yu H.
  TITLE     pUMRI plasmids for metabolic pathway assembly
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5901)
  AUTHORS   Xie W, Lv X, Yu H.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JUL-2014) Department of Chemical and Biological 
            Engineering, Zhejiang University, Institute of Bioengineering, 34 
            TianMuShan Road, Hangzhou, Zhejiang 310028, China
REFERENCE   3  (bases 1 to 5901)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5901)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-JUL-2014) Department of Chemical and Biological Engineering, 
            Zhejiang University, Institute of Bioengineering, 34 TianMuShan 
            Road, Hangzhou, Zhejiang 310028, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Artificial Sequence v. Artificial Sequence 
            Sequencing Technology :: Artificial Sequence
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..55
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    88..1095
                     /label=GAL1-7 homologous arm
                     /note="GAL1-7 homologous arm"
     terminator      complement(1106..1271)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1419..1442)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        1489..2153
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        2164..2182
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2200..2229
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      2259..2448
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2461..2479)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(2502..2535)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2581..3937
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        4032..4252
                     /label=URA3 promoter
     CDS             4253..5053
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5225..5813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggatcttcga gaacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt
       61 gatatcagat ccacggatca ctaaagggaa caaaagctgg agctggcctt gttggcaagc
      121 attgactgac gatctctcca gagaagcaag agaaaatcat aagcctttca aatatgtcca
      181 aatatttgaa aacaaaggta cagccatggg ttgttccaac ttacatccac atggccaagc
      241 ttggtgctta gaatccatcc ctagtgaagt ttcgcaagaa ttgaaatctt ttgataaata
      301 taaacgtgaa cacaatactg atttgtttgc cgattacgtc aaattagaat caagagagaa
      361 gtcaagagtc gtagtggaga atgaatcctt tattgttgtt gttccatact gggccatctg
      421 gccatttgag accttggtca tttcaaagaa gaagcttgcc tcaattagcc aatttaacca
      481 aatggtgaag gaggacctcg cctcgatttt aaagcaacta actattaagt atgataattt
      541 atttgaaacg agtttcccat actcaatggg tatccatcag gccataaagg ccgaagaaga
      601 caatccactg ccagttggta catcaccctc acagaagact tgcagcccgg ccagaggagc
      661 actggcaaac ctttccggtg caagtttctt tagaaaagag tgagcaacat ggagaccaca
      721 tttaaagtaa ttagaccagt ccgacacaga aggatcaatt gtgacataag aaccgtccaa
      781 cggcaaatcg aacttccttt gagcaaattt gggatcagca tttattaagg taatggatgg
      841 atttttctcg ttcaaaactt tgacggcgca aagcatatca aaatcaatag ctaaaggtaa
      901 aaccgagaag tcacaataat caatatgttc accaattaga ttgactctac caggcgatct
      961 agcaacaaaa tccggtttag catcataagc gcttataaat ttcttaatta tgctcgggca
     1021 cttttcggcc aatggtcttg gtaattcctt tgcgctagaa ttgaactcag gtgtaggcct
     1081 cttcgctatt acgccagctg aattggagcg acctcatgct atacctgaga aagcaacctg
     1141 acctacagga aagagttact caagaataag aattttcgtt ttaaaaccta agagtcactt
     1201 taaaatttgt atacacttat tttttttata acttatttaa taataaaaat cataaatcat
     1261 aagaaattcg cttatttaga agtgtcaaca acgtatctac caacgatttg acccttttcc
     1321 atcttttcgt aaatttctgg caaggtagac aagccgacaa ccttgattgg agacttgacc
     1381 aaacctctgg cgaagaattg ttaattaaga gctcagatct tatcgtcgtc atccttgtaa
     1441 tccatcgata ctagtgcggc cgccctttag tgagggttga attcgaattt tcaaaaattc
     1501 ttactttttt tttggatgga cgcaaagaag tttaataatc atattacatg gcattaccac
     1561 catatacata tccatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa
     1621 agagccccat tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta
     1681 actgctcatt gctatattga agtacggatt agaagccgcc gagcgggtga cagccctccg
     1741 aaggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
     1801 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
     1861 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta
     1921 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
     1981 cgaagcgatg atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt
     2041 taactaatac tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta
     2101 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat
     2161 ccgtaatacg actcactata gggcccgggc gtcgacatgg aacagaagtt gatttccgaa
     2221 gaagacctcg agtaagcttg gtaccgcggc tagctaagat ccgctctaac cgaaaaggaa
     2281 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     2341 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     2401 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaagat cccaattcgc
     2461 cctatagtga gtcgtattac gcgcgctcga caacccttaa tataacttcg tataatgtat
     2521 gctatacgaa gttattaggt ctagtagctt gcctcgtccc cgccgggtca cccggccagc
     2581 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
     2641 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
     2701 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
     2761 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
     2821 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
     2881 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga
     2941 ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     3001 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     3061 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     3121 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     3181 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc
     3241 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     3301 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     3361 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     3421 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
     3481 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     3541 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     3601 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3661 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     3721 atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     3781 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     3841 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     3901 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     3961 ccatccagtg tcgaaaacga acagtgtggc tgtggttcag attgtactga gagtgcacca
     4021 taccacagct tttcaattca attcatcatt ttttttttat tctttttttt gatttcggtt
     4081 tctttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     4141 cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
     4201 attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa tcatgtcgaa
     4261 agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa
     4321 tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga
     4381 attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga
     4441 tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa
     4501 gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata cagtcaaatt
     4561 gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg
     4621 tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga
     4681 acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga
     4741 atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat
     4801 tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg
     4861 tgtgggttta gatgacaagg gagacgcatt gggtcaacag tatagaaccg tggatgatgt
     4921 ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga
     4981 tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg
     5041 cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat
     5101 tagagcttca atttaattat atcagttatt accctatgcg gtgtgaaata ccgcacagat
     5161 gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt atttctataa tgattgctgg
     5221 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5281 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5341 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5401 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5461 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5521 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5581 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5641 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5701 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5761 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5821 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5881 tggtcatgag attatcaaaa a
//
