LOCUS       40924_32295        5057 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-40, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  AUTHORS   Xie W, Lv X, Yu H.
  TITLE     pUMRI plasmids for metabolic pathway assembly
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5057)
  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 5057)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5057)
  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            "Unpublished"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..5057
                     /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    113..917
                     /label=LPP1 homologous arm
                     /note="LPP1 homologous arm"
     terminator      complement(947..1217)
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate
                     kinase gene"
     CDS             complement(1236..1259)
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     regulatory      complement(1293..1811)
                     /label=GAL2 promoter from Saccharomyces cerevisiae
                     /note="GAL2 promoter from Saccharomyces cerevisiae"
                     /regulatory_class="promoter"
     regulatory      1812..2360
                     /label=GAL7 promoter from Saccharomyces cerevisiae
                     /note="GAL7 promoter from Saccharomyces cerevisiae"
                     /regulatory_class="promoter"
     promoter        2368..2386
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2404..2433
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     regulatory      2459..2791
                     /label=TPS1 terminator from Saccharomyces cerevisiae
                     /note="TPS1 terminator from Saccharomyces cerevisiae"
                     /regulatory_class="terminator"
     misc_feature    complement(2813..2846)
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2813..2846)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2892..4248
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(4381..4969)
                     /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 gtaaattctt
      121 acgcgtgaac caaaacaaaa attatcaacc aatacatgtc gacaaataat gaattagagc
      181 aatagccacg ataacagacc ttgattaaat gtaaactaca tataagttta taagggttgg
      241 ttgattccat tattttttgt tttttatata aagcctatga aaaataaatg gaagcggctg
      301 ttagaatata aaatagaagt tgaaacaaca aaaatgtaag aaagttgcca acaataaatc
      361 tcaaacttga aagtgtggat cgaggaatat ggacaatctt taaaaaattt ttagtcccgt
      421 agtttttcga atttttcctt accagtgacc atgaaaccat gattggccat aaaggccatc
      481 ggtattttgg cttcggttaa tatctggcgt ggacccatcg gtggaaaaac gccacagttt
      541 tcaagctcaa ttatagtcac gtgggtagaa atattacact gcaccgcaag aaagtacgac
      601 cagatgaatc acatgtgaag atgttaacaa gtttacatac tttttgatca ggtaagctat
      661 gcataatgtc ttagttttag ttatgtaaag aaatgcaaac ttttagtgca tattttatct
      721 tatataatag agcaaagctc gccagtgctc cagttgttac ctgagaatga agcttgatcg
      781 gcttttattc tttcctgata ggactctgta agtgttacta aacactaacc ggtgaaggaa
      841 gtatgtctct tttcgccaag tttgtaaatg tccatttcca gcaacaataa atgactaaaa
      901 atgctagaac agctccatgt aggcctcttc gctattacgc ctaacgaacg cagaattttc
      961 gagttattaa acttaaaata cgctgaaccc gaacatagaa atatcgaatg ggaaaaaaaa
     1021 actgcataaa ggcattaaaa gaggagcgaa ttttttttta ataaaaatct taataatcat
     1081 taaaagataa ataatagtct atatatacgt atataaataa aaaatattca aaaaataaaa
     1141 taaactatta ttttagcgta aaggatgggg aaagagaaaa gaaaaaaatt gatctatcga
     1201 tttcaattca attcaattta attaagagct cagatcttat cgtcgtcatc cttgtaatcc
     1261 atcgatacta gtgcggccgc agggttgaat tctatgaaag aattattttt tttattatgt
     1321 taatcttgtg tttacttaac tattactatt cttgatgata attgaataag gtgcataatg
     1381 aagagcaatt cacaacacca aattttcaat ccaattactg attgtttata tatgtctaca
     1441 aaacttatcc tatctccaca ttttagcctg cgaaatgttt gttttttgaa caatagctct
     1501 ccagaatgtt gtataattta agaatatgtg cacagttaac tttctagcag gagtataatg
     1561 ccatttgctc cccatcttga gatgggaagg gcttaactaa tctcggttcg gagtgatccg
     1621 ccccgatact gccttctgcc ttaatatcgt ccaaggcaca tggacccctg aacggcgcag
     1681 atatctccgc acggacgaaa gaccgccggt gccttcctga ggcaaccgcc cctttcgaat
     1741 atagatcacg tgacccattt ttagctacta atagaaaaag aaattgcaac ctacttaagc
     1801 cattccggaa gctgtaccac catggagaca tcaaaaattg aaaatctatg gaaagatatg
     1861 gacggtagca acaagaatat agcacgagcc gcggagttca tttcgttact tttgatatca
     1921 ctcacaacta ttgcgaagcg cttcagtgaa aaaatcataa ggaaaagttg taaatattat
     1981 tggtagtatt cgtttggtaa agtagagggg gtaatttttc ccctttattt tgttcataca
     2041 ttcttaaatt gctttgcctc tccttttgga aagctatact tcggagcact gttgagcgaa
     2101 ggctcattag atatattttc tgtcattttc cttaacccaa aaataaggga aagggtccaa
     2161 aaagcgctcg gacaactgtt gaccgtgatc cgaaggactg gctatacagt gttcacaaaa
     2221 tagccaagct gaaaataatg tgtagctatg ttcagttagt ttggctagca aagatataaa
     2281 agcaggtcgg aaatatttat gggcattatt atgcagagca tcaacatgat aaaaaaaaac
     2341 agttgaatat tccctcaaaa ggatccgtaa tacgactcac tatagggccc gggcgtcgac
     2401 atggaacaga agttgatttc cgaagaagac ctcgagtaag cttggtaccg cggctagctg
     2461 aacccgatgc aaatgagacg atcgtctatt cctggtccgg ttttctctgc cctctcttct
     2521 attcactttt tttatacttt atataaaatt atataaatga cataactgaa acgccacacg
     2581 tcctctccta ttcgttaacg cctgtctgta gcgctgttac tgaagctgcg caagtagttt
     2641 tttcaccgta taggccctct ttttctctct ctttctttct ctcccgcgct gatctcttct
     2701 tcgaaacatc atgaataaaa agaaaaagga aatcaagaaa aaaaagccat aatttatccc
     2761 acattttttt ttattgtcgc tgttcacacc ggcgcgctcg acaaccctta atataacttc
     2821 gtataatgta tgctatacga agttattagg tctagtagct tgcctcgtcc ccgccgggtc
     2881 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
     2941 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
     3001 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
     3061 agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
     3121 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
     3181 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg
     3241 taaggaaaag actcacgttt cgaggccgcg attaaattcc aacatggatg ctgatttata
     3301 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     3361 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     3421 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     3481 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggcaa
     3541 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3601 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3661 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3721 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3781 gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3841 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3901 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3961 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     4021 tttgatgctc gatgagtttt tctaatcagt actgacaata aaaagattct tgttttcaag
     4081 aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc
     4141 gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag
     4201 aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga
     4261 tactaacgcc gccatccagt gtcgaaaacg aacagaatca ggggataacg caggaaagaa
     4321 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4381 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4441 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4501 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4561 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4621 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4681 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4741 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4801 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4861 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4921 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4981 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5041 catgagatta tcaaaaa
//
