LOCUS       40924_32280        5028 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMRI-35, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5028)
  AUTHORS   Xie W, Yu H.
  TITLE     Developing a sequential repressing-inducing control strategy for 
            improving carotenoids production in Saccharomyces cerevisiae
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5028)
  AUTHORS   Xie W, Yu H.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2014) Department of Chemical and Biological 
            Engineering, Zhejiang University, Institute of Bioengineering, 34 
            TianMuShan Road, Hangzhou, Zhejiang 310028, China
REFERENCE   3  (bases 1 to 5028)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5028)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-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"
FEATURES             Location/Qualifiers
     source          1..5028
                     /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"
     promoter        1911..2318
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2339..2357
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2375..2404
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     regulatory      2430..2762
                     /label=TPS1 terminator derived from S. cerevisiae
                     /note="TPS1 terminator derived from S. cerevisiae"
                     /regulatory_class="terminator"
     misc_feature    2784..2817
                     /label=Loxp site
                     /note="Loxp site"
     protein_bind    complement(2784..2817)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            2863..4219
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(4352..4940)
                     /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 tttttgatta aaattaaaaa aactttttgt ttttgtgttt
     1321 attctttgtt cttagaaaag acaagttgag cttgtttgtt cttgatgttt tattatttta
     1381 caatagctgc aaatgaagaa tagattcgaa cattgtgaag tattggcata tatcgtctct
     1441 atttatactt tttttttttc agttctagta tattttgtat tttcctcctt ttcattcttt
     1501 cagttgccaa taagttacag gggatctcga aagatggtgg ggatttttcc ttgaaagacg
     1561 actttttgcc atctaatttt tccttgttgc ctctgaaaat tatccagcag aagcaaatgt
     1621 aaaagatgaa cctcagaaga acacgcaggg gcccgaaatt gttcctacga gaagtagtgg
     1681 gtcataaaaa gtttattccc tggaaaaaaa attttgcgtt gcctttctgg agaatttttt
     1741 cgaattagcg tgctgccact gcatgcattt ctgagaagtg tgggcattct tccaccagtt
     1801 gttcctccta aaaaaaaaaa agatttccta ccccgcacaa attcctgcat acccctcatt
     1861 tccacgaccg cgaatcctta catcacaccc aatcccccac aagtgatccc ccacacacca
     1921 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
     1981 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
     2041 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
     2101 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     2161 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
     2221 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
     2281 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa aggatccgta
     2341 atacgactca ctatagggcc cgggcgtcga catggaacag aagttgattt ccgaagaaga
     2401 cctcgagtaa gcttggtacc gcggctagct gaacccgatg caaatgagac gatcgtctat
     2461 tcctggtccg gttttctctg ccctctcttc tattcacttt ttttatactt tatataaaat
     2521 tatataaatg acataactga aacgccacac gtcctctcct attcgttaac gcctgtctgt
     2581 agcgctgtta ctgaagctgc gcaagtagtt ttttcaccgt ataggccctc tttttctctc
     2641 tctttctttc tctcccgcgc tgatctcttc ttcgaaacat catgaataaa aagaaaaagg
     2701 aaatcaagaa aaaaaagcca taatttatcc cacatttttt tttattgtcg ctgttcacac
     2761 cggcgcgctc gacaaccctt aatataactt cgtataatgt atgctatacg aagttattag
     2821 gtctagtagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
     2881 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
     2941 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
     3001 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
     3061 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
     3121 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
     3181 cacatcacat ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc
     3241 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     3301 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     3361 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     3421 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     3481 catggttact caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc
     3541 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     3601 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     3661 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     3721 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     3781 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     3841 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     3901 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     3961 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     4021 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     4081 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     4141 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     4201 aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag tgtcgaaaac
     4261 gaacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaa
//
