LOCUS       40924_33317        6352 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNK071, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6352)
  AUTHORS   Kandul NP, Guo M, Hay BA.
  TITLE     A positive readout single transcript reporter for site-specific mRNA
            cleavage
  JOURNAL   PeerJ (2017) In press
REFERENCE   2  (bases 1 to 6352)
  AUTHORS   Kandul NP, Guo M, Hay BA.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JAN-2017) Biology and Biological Engineering, 
            California Institute of Technology, 1200 E. California Blvd., 
            Pasadena, CA 91125, USA
REFERENCE   3  (bases 1 to 6352)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6352)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PeerJ 
            (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-JAN-2017) Biology and Biological Engineering, California 
            Institute of Technology, 1200 E. California Blvd., Pasadena, CA 
            91125, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2606..3301
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     intron          3319..3598
     ncRNA           3337..3548
                     /label=miRNA
                     /note="miR-6.1 backbone with a synthetic guide miRNA"
                     /ncRNA_class="miRNA"
     ncRNA           3458..3479
                     /label=other
                     /note="synthetic guide miRNA"
                     /ncRNA_class="other"
     CDS             3637..3678
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             3688..3705
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    complement(3778..3912)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4534..5122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5296..6153)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6154..6258)
                     /label=AmpR promoter
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 ctactagtcc agtgtggtgg aattcatggt cagcgagctg atcaccgaga acatgcacat
     2641 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
     2701 aggcaagccc tacgagggca cccagaccat gagaatcaag gctgtcgagg gcggccctct
     2761 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa
     2821 ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga
     2881 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     2941 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc
     3001 tgtgatgcag aagaaaacac tcggctggga ggccagcacc gagacactgt accccgctga
     3061 cggcggcctg gaaggcagag cggacatggc cctgaagctc gtgggcgggg gccacctgat
     3121 ctgcaacttg aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg
     3181 cgtctactat gtggacagaa gactggaaag aatcaaggag gccgacaaag agacctacgt
     3241 cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcaccg
     3301 gcttaattag gctagcaggt aagtattaac tatttattta aagtccacaa ctcatcaagg
     3361 aaaatgaaag tcaaagttgg cagcttactt aaacttaatc acagccttta atgtccactc
     3421 ctcgagttcg aaccattaag ttaatatacc atatctaatt gttcgaactc gaggagtggg
     3481 tacctaaagt gcctaacatc attatttaat ttttattttt attggcacac gaataaccat
     3541 gccgttttgc aagtactaat ccttattaat aacttttttc tttttccttt cttttcagcg
     3601 tgcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
     3661 ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc
     3721 tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc gatcgaggat
     3781 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3841 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3901 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3961 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag
     4021 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     4081 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     4141 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
     4201 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
     4261 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
     4321 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
     4381 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
     4441 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc agcaaaaggc
     4501 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4561 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4621 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4681 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     4741 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4801 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4861 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4921 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     4981 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5041 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5101 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5161 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5221 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5401 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5581 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     5641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5881 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     5941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6241 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6301 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gt
//
