LOCUS       40924_13370        4881 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCrCre DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4881)
  AUTHORS   Kasai Y, Harayama S.
  TITLE     Construction of marker-free transgenic strains of Chlamydomonas 
            reinhardtii using a Cre/loxP-mediated recombinase system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4881)
  AUTHORS   Kasai Y, Harayama S.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2016) Contact:Yuki Kasai Chuo University, 
            Department of Biological sciences; Kasuga 1-13-27, Bunkyo-ku, Tokyo 
            112-8551, Japan
REFERENCE   3  (bases 1 to 4881)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4881)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAY-2016) Contact:Yuki Kasai Chuo University, Department of 
            Biological sciences; Kasuga 1-13-27, Bunkyo-ku, Tokyo 112-8551, 
            Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..34
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      67..333
                     /note="includes promoter region of Chlamydomonas
                     reinhardtii nuclear gene hsp70A for cytosolic HSP70"
                     /regulatory_class="promoter"
     regulatory      342..574
                     /note="includes promoter region and start codon of
                     Chlamydomonas reinhardtii nuclear gene rbcS2 for ribulose 
                     bisphosphate carboxylase/oxygenase small subunit"
                     /regulatory_class="promoter"
     gene            567..574
                     /gene="CrCRE"
                     /label=CrCRE
     intron          575..719
                     /note="includes intron 1 from Chlamydomonas reinhardtii
                     rbcS2 gene"
     CDS             727..1755
                     /label=Cre
                     /note="site-specific recombinase"
     regulatory      1759..1998
                     /note="includes terminator region from Chlamydomonas
                     reinhardtii rbcS2 gene"
                     /regulatory_class="terminator"
     promoter        complement(2037..2055)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2065..2081)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2222..2677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2704..2808
                     /label=AmpR promoter
     CDS             2809..3666
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3840..4428
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4716..4737
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4752..4782
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4790..4806
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4814..4830
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4851..4869
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag
       61 cccggggagc tcgctgaggc ttgacatgat tggtgcgtat gtttgtatga agctacagga
      121 ctgatttggc gggctatgag ggcgggggaa gctctggaag ggccgcgatg gggcgcgcgg
      181 cgtccagaag gcgccatacg gcccgctggc ggcacccatc cggtataaaa gcccgcgacc
      241 ccgaacggtg acctccactt tcagcgacaa acgagcactt atacatacgc gactattctg
      301 ccgctataca taaccactca gctagcttaa gatcccatca agcttgcatg ccgggcgcgc
      361 cagaaggagc gcagccaaac caggatgatg tttgatgggg tatttgagca cttgcaaccc
      421 ttatccggaa gccccctggc ccacaaaggc taggcgccaa tgcaagcagt tcgcatgcag
      481 cccctggagc ggtgccctcc tgataaaccg gccagggggc ctatgttctt tactttttta
      541 caagagaagt cactcaacat cttaaaatgg ccaggtgagt cgacgagcaa gcccggcgga
      601 tcaggcagcg tgcttgcaga tttgacttgc aacgcccgca ttgtgtcgac gaaggctttt
      661 ggctcctctg tcgctgtctc aagcagcatc taaccctgcg tcgccgtttc catttgcagg
      721 atggccatga gcaacctgct gaccgtgcac cagaacctgc ccgccctgcc cgtggacgcg
      781 acgagcgacg aggtgcgcaa gaacctgatg gacatgttcc gcgaccgcca ggccttctcg
      841 gagcacacct ggaagatgct gctgagcgtg tgccgctcgt gggccgcgtg gtgcaagctg
      901 aacaaccgca agtggttccc cgcggagccg gaggacgtgc gcgactacct gctgtacctg
      961 caggcccgcg gcctggcggt gaagacgatc cagcagcacc tgggccagct gaacatgctg
     1021 caccgccgca gcggcctgcc ccgcccgagc gactcgaacg ccgtgtcgct ggtcatgcgc
     1081 cgcatccgca aggagaacgt ggacgccggc gagcgcgcga agcaggccct ggcgttcgag
     1141 cgcaccgact tcgaccaggt gcgctccctg atggagaaca gcgaccgctg ccaggacatc
     1201 cgcaacctgg ccttcctggg catcgcgtac aacaccctgc tgcgcatcgc cgagatcgcg
     1261 cgcatccgcg tgaaggacat cagccgcacg gacggcggcc gcatgctgat ccacatcggc
     1321 cgcaccaaga cgctggtgtc gaccgccggc gtggagaagg cgctgtccct gggcgtgacg
     1381 aagctggtgg agcgctggat ctccgtgagc ggcgtggccg acgaccccaa caactacctg
     1441 ttctgccgcg tgcgcaagaa cggcgtggcc gcgccgtcgg cgacctccca gctgagcacg
     1501 cgcgccctgg agggcatctt cgaggcgacc caccgcctga tctacggcgc caaggacgac
     1561 tccggccagc gctacctggc ctggtcgggc cactccgcgc gcgtgggcgc cgcgcgcgac
     1621 atggcccgcg cgggcgtgag catccccgag atcatgcagg ccggcggctg gaccaacgtg
     1681 aacatcgtga tgaactacat ccgcaacctg gactccgaga cgggcgcgat ggtgcgcctg
     1741 ctggaggacg gcgactaagg atccccgctc cgtgtaaatg gaggcgctcg ttgatctgag
     1801 ccttgccccc tgacgaacgg cggtggatgg aagatactgc tctcaagtgc tgaagcggta
     1861 gcttagctcc ccgtttcgtg ctgatcagtc tttttcaaca cgtaaaaagc ggaggagttt
     1921 tgcaattttg ttggttgtaa cgatcctccg ttgattttgg cctctttctc catgggcggg
     1981 ctgggcgtat ttgaagcgtc tagagcggcc gccaccgcgg tggagctcca attcgcccta
     2041 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     2101 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     2161 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2221 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2281 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2341 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2401 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2461 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2521 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2581 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2641 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2701 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2761 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2821 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2881 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2941 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3001 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3061 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3121 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3181 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3241 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3301 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3361 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3421 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3481 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3541 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3601 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3661 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3721 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3781 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3841 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3901 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3961 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4021 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4081 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4141 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4201 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4261 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4321 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4381 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4441 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4501 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4561 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4621 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4681 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     4741 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     4801 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     4861 cactaaaggg aacaaaagct g
//
