LOCUS       62056_13780        7105 bp DNA     circular SYN 21-APR-2021
DEFINITION  Cloning vector pleCC, complete sequence.
ACCESSION   MW770351
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7105)
  AUTHORS   Li Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAR-2021) Cell and Developmental Biology, University 
            of California, San Diego, 9500 Gilman Drive Tata Hall 5401, San 
            Diego, CA 92122, USA
REFERENCE   2  (bases 1 to 7105)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (18-MAR-2021) Cell and Developmental Biology, University of 
            California, San Diego, 9500 Gilman Drive Tata Hall 5401, San Diego, 
            CA 92122, USA"
FEATURES             Location/Qualifiers
     source          1..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(127..715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(889..1746)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1747..1851)
                     /label=AmpR promoter
     CDS             3223..3897
                     /label=DsRed2
                     /note="improved tetrameric variant of DsRed fluorescent
                     protein"
     CDS             3907..3927
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    4068..4189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4192..4612
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3
                     snRNA (Port et al., 2014)"
     ncRNA           4612..4630
                     /label=guide_RNA
                     /note="ple Intron gRNA"
                     /ncRNA_class="guide_RNA"
     misc_RNA        4632..4707
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      4714..4941
                     /label=U6 terminal
                     /note="U6 terminal"
                     /regulatory_class="terminator"
     polyA_signal    complement(5007..5136)
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     CDS             complement(5154..5870)
                     /label=mCerulean
                     /note="enhanced monomeric variant of CFP (Rizzo et al.,
                     2004)"
     regulatory      6048..6091
                     /label=3XP3
                     /note="3XP3"
                     /regulatory_class="promoter"
ORIGIN
        1 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
       61 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      121 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      181 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      241 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      301 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      361 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      421 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      481 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      541 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      601 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      661 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
      961 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1081 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1201 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1321 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1381 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1501 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1561 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1741 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1801 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1861 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     1921 taggcgtatc acgaggccct ttcgttgttc accccaaacg aaaacaaccc aaacacacaa
     1981 aaaaaaatga tggccgttgc agcagcccaa aagaaccgcg agatgttcgc catcaagaaa
     2041 tcctacagta ttgaggtgag tcgaaaatcc agcaaaacat acaatttcca aatcgaattg
     2101 taaaaaaaaa gaagcaaatg agtgactaga tttggtctac gtgactatct aaaggttttg
     2161 tgttttcaaa aaaaaaccag ttaaccggcc agtagtcgag cccatttttg tgatcccgca
     2221 tttctttttc gccaattcaa tttgaccaaa ctcaatcctc ggttggccat taagtcacac
     2281 ctctaagaac aggcggccca aatcgaaagg cccaaatcgc caacggacac aacgggttgt
     2341 gtgttaagtt attatttaac aaataagtga aagtggctgc ctttgggccc catgcactta
     2401 gcctggccga aacagtcgcg cctcattgtt gttgactcgt ctctgtttat gtttatgctg
     2461 cgtccgagca aaacaaaaaa tatgaaatca atataaatga ttcaatgagt tggtcatctt
     2521 tcatcagcca aagcggtggc ccaaaaaaaa aaactgaaga aaacaaacaa acaaaaaaaa
     2581 aaaaaaacga gctgacagtg aaaactgcaa gtgatggaaa cctttttccc cccatcaaga
     2641 aaaagaaaga aaaaaaacct ttaacgatcg ccagttgcat ttttttcccg ccagaatcgc
     2701 atggggattt tcgatttctc agtcacatat catttatctt cgcacctgcc cgcttcgaag
     2761 cggcgacatc gcattaaatt accattcgtg gctttgaatc acaatcaata cagttccatg
     2821 aatagccagc tttattcaaa gaaaaagatc tgaagacggc gctttcacac accattttat
     2881 ataatatttt caaaattgcg caatactctg ctcaaagtca ccacccaagc gtggagccga
     2941 agaaagccac cctggcttga aggtcaaggc aatcgtctaa tttccgctcc acatcctgcc
     3001 agtggtccat catgatggtc gactctagaa gtcgatccaa catggcgact tgtcccatcc
     3061 ccggcatgtt taaatatact aattattctt gaactaattt taatcaaccg atttatctct
     3121 cttccgcagg tgggaggttc cggtggaagc ggaggtagcg gcggatccga gggccgcggc
     3181 agcctgctga cctgcggcga tgtggaggag aaccccgggc ccatggcctc ctccgagaac
     3241 gtcatcaccg agttcatgcg cttcaaggtg cgcatggagg gcaccgtgaa cggccacgag
     3301 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gccacaacac cgtgaagctg
     3361 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttccag
     3421 tacggctcca aggtgtacgt gaagcacccc gccgacatcc ccgactacaa gaagctgtcc
     3481 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggcgacc
     3541 gtgacccagg actcctccct gcaggacggc tgcttcatct acaaggtgaa gttcatcggc
     3601 gtgaacttcc cctccgacgg ccccgtgatg cagaagaaga ccatgggctg ggaggcctcc
     3661 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agacccacaa ggccctgaag
     3721 ctgaaggacg gcggccacta cctggtggag ttcaagtcca tctacatggc caagaagccc
     3781 gtgcagctgc ccggctacta ctacgtggac gccaagctgg acatcacctc ccacaacgag
     3841 gactacacca tcgtggagca gtacgagcgc accgagggcc gccaccacct gttcctgaga
     3901 tctcgaccca agaaaaagcg gaaggtggag gacccgtaag atccaccgga tctagataac
     3961 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4021 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4081 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4141 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttac cttttttgct
     4201 cacctgtgat tgctcctact caaatacaaa aacatcaaat tttctgtcaa taaagcatat
     4261 ttatttatat ttattttaca ggaaagaatt ccttttaaag tgtattttaa cctataatga
     4321 aaaacgatta aaaaaaatac ataaaataat tcgaaaattt ttgaatagcc caggttgata
     4381 aaaattcatt tcatacgttt tataacttat gcccctaagt attttttgac catagtgttt
     4441 caattctaca ttaattttac agagtagaat gaaacgccac ctactcagcc aagaggcgaa
     4501 aaggttagct cgccaagcag agagggcgcc agtgctcact actttttata attctcaact
     4561 tctttttcca gactcagttc gtatatatag acctattttc aatttaacgt cgaagtccgt
     4621 gttagtgaca cgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     4681 cttgaaaaag tggcaccgag tcggtgcttt tttgcctacc tggagcctga gagttgttca
     4741 ataaaataaa aatgtttcgt ttttttgctt tcgccagtat ttattatttt tcatcaatat
     4801 gtattcaatt tggtatgtat ttagtaattg taatatatag acaatggttt tccgttgacg
     4861 tacatacatc tgacgtgtgt ttatttagac ataatagtta tgttttcaca tctttttaat
     4921 gttcgcttaa tgcgtatgca tttagaccta cgcccccaac tgagagaact caaaggttac
     4981 cccagttggg gcactacgat cgatcccacg cgcttgaaag gagtgtgtaa atggacaatg
     5041 ttgttacaat aaaagttgaa attattttat aaaatttatt aaaaacatat ttaaacataa
     5101 catattgtat taaaacatga caatacaaac taagatttag tcagataaac ctatttgtag
     5161 agttcgtcca tacccaaagt aattcctgcc gcggtcacga actccagcaa caccatatga
     5221 tccctctttt cgttgggatc cttactgagt ttactctgcg tgctcaggta gtggttgtca
     5281 ggcaagagca cgggaccgtc gccgataggt gtattttgct gataatgatc ggccaattgg
     5341 accgatccat cttctatgtt acaattcaga ccgaaattcg ctttgatacc gtttttctgt
     5401 ttatcggctg ttatataaac atttccatgg atagcgttat attccaactt atggcccaga
     5461 atattaccat cttccttaaa gtctataccc ttcaactcga tacgattcac cagtgtatca
     5521 ccttcaaact tgacctctgc ccgtgttttg tagttgccat cgtctttgaa aaatattgtc
     5581 cgttcctgca cataaccttc tggcattgca cttttaaaga agtcgtgctg cttcatgtgg
     5641 tccgggtaac gagcgaaaca ctgaactccc cacgagagtg tagtcacgag tgtcggccag
     5701 ggtacaggca gtttgcccgt ggtgcaaatg aatttcaggg tcagtttacc gtaagtagca
     5761 tctccttcac cttctccact gacactgaat ttgtgaccgt tcacgtcacc atccaattcg
     5821 acgagtatcg ggaccacgcc agtgaagagt tcttcacctt tactcaccat ggtggcgacc
     5881 ggcttcgagc cgattgttta gcttgttcag ctgcgcttgt ttatttgctt agctttcgct
     5941 tagcgacgtg ttcactttgc ttgtttgaat tgaattgtcg ctccgtagac gaagcgcctc
     6001 tatttatact ccggcggtcg agggttcgaa atcgataagc ttggatccta attgaattag
     6061 ctctaattga attagtctct aattgaatta gatccccggg cgagctcgcc taggtcacta
     6121 acacggactt caatcttttc gctttcgttt ccagaatggc tatccatccc gccgtcgcag
     6181 cctggtggat gatgcccgtt tcgagaccct ggtggtcaag cagaccaaac aaaccgtcct
     6241 cgaggaggcc cgcagcaagg caaatggtaa gtggataggg tccaacaagt gggtcactaa
     6301 aatagctcca aagagggttt ctatttggga tcaaaaggga aattttgcag ctagggcttt
     6361 acttgttgat taaaaactta aaaagttgca atttttattt cattttattc gttttgacat
     6421 agatttgcgt ttttttctta ttcattttaa ccctttattt attatttgcc ttcgttttta
     6481 tttttatgat tttaaagacg attccctgga agattgcatt gtgcaggctc aggagcacat
     6541 tccctccgag caagatgtgg agctccagga cgagcatgcc aatctggaga acctgccctt
     6601 ggaggaatat gttccaggtt aatatccatt tccatttcaa ttcccatgcc agttactcct
     6661 ctttctgagc ctattatctt tgatttgatt ttgattatga ttttcatggg ctttattatt
     6721 atttttattt cattcgcttt tagttgagga ggatgttgag tttgagagcg ttgaacagga
     6781 gcagtcggag tcgcagtcgc aggagccaga aggcaaccaa cagccgacca agaacggtga
     6841 gcagggggtt ctagaatcca gcagccaacc ataagtagta cttaacttct gagaggggct
     6901 tttgaaaagc gtgcgacatt gatttaagtg cttaatatga taaagtgcca gggcagagga
     6961 aaagcactca gccacacgga caattgtgag tgggcgcgct cgctcgctct ctctctctct
     7021 ctctctctct ctgtggaaat gtcgcatttc cctttttcat ttgttacgct cgacttaata
     7081 caagtacgaa atgcgtttgc ctggc
//
