LOCUS       40924_45753        4939 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUMRI-21, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4939)
  AUTHORS   Xie W, Lv X, Yu H.
  TITLE     pUMRI plasmids for metabolic pathway assembly
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4939)
  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 4939)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4939)
  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            "Unpublished"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..4939
                     /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)."
     terminator      complement(144..309)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(457..480)
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     promoter        527..1191
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     promoter        1202..1220
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1238..1267
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      1297..1486
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1499..1517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(1540..1573)
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(1540..1573)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            1619..2975
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        3070..3290
                     /label=URA3 promoter
     CDS             3291..4091
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4263..4851)
                     /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 gtaggcctct
      121 tcgctattac gccagctgaa ttggagcgac ctcatgctat acctgagaaa gcaacctgac
      181 ctacaggaaa gagttactca agaataagaa ttttcgtttt aaaacctaag agtcacttta
      241 aaatttgtat acacttattt tttttataac ttatttaata ataaaaatca taaatcataa
      301 gaaattcgct tatttagaag tgtcaacaac gtatctacca acgatttgac ccttttccat
      361 cttttcgtaa atttctggca aggtagacaa gccgacaacc ttgattggag acttgaccaa
      421 acctctggcg aagaattgtt aattaagagc tcagatctta tcgtcgtcat ccttgtaatc
      481 catcgatact agtgcggccg ccctttagtg agggttgaat tcgaattttc aaaaattctt
      541 actttttttt tggatggacg caaagaagtt taataatcat attacatggc attaccacca
      601 tatacatatc catatacata tccatatcta atcttactta tatgttgtgg aaatgtaaag
      661 agccccatta tcttagccta aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac
      721 tgctcattgc tatattgaag tacggattag aagccgccga gcgggtgaca gccctccgaa
      781 ggaagactct cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg
      841 cctcgcgccg cactgctccg aacaataaag attctacaat actagctttt atggttatga
      901 agaggaaaaa ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac
      961 aaccatagga tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg
     1021 aagcgatgat ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta
     1081 actaatactt tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc
     1141 aacaaaaaat tgttaatata cctctatact ttaacgtcaa ggagaaaaaa ccccggatcc
     1201 gtaatacgac tcactatagg gcccgggcgt cgacatggaa cagaagttga tttccgaaga
     1261 agacctcgag taagcttggt accgcggcta gctaagatcc gctctaaccg aaaaggaagg
     1321 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     1381 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     1441 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaagatcc caattcgccc
     1501 tatagtgagt cgtattacgc gcgctcgaca acccttaata taacttcgta taatgtatgc
     1561 tatacgaagt tattaggtct agtagcttgc ctcgtccccg ccgggtcacc cggccagcga
     1621 catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg
     1681 tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg catccataca
     1741 ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca
     1801 gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa
     1861 tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt
     1921 gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa ggaaaagact
     1981 cacgtttcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2041 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     2101 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2161 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2221 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggcaaaac agcattccag
     2281 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2341 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2401 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     2461 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc
     2521 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2581 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2641 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2701 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2761 gagtttttct aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg
     2821 tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt
     2881 tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
     2941 gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc
     3001 atccagtgtc gaaaacgaac agtgtggctg tggttcagat tgtactgaga gtgcaccata
     3061 ccacagcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
     3121 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     3181 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     3241 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
     3301 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     3361 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     3421 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     3481 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     3541 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     3601 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     3661 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     3721 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     3781 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     3841 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     3901 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     3961 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     4021 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     4081 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     4141 gagcttcaat ttaattatat cagttattac cctatgcggt gtgaaatacc gcacagatgc
     4201 gtaaggagaa aataccgcat caggaaattg taaacgttat ttctataatg attgctggcg
     4261 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4321 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4381 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4441 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4501 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4561 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4621 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4681 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4741 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4801 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4861 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4921 gtcatgagat tatcaaaaa
//
