LOCUS       Exported                4440 bp ds-DNA     circular SYN 12-AUG-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    PV521
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Aug 12, 2019 from SnapGene 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4440
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        103..1102
                     /label=ZM-U6 POLIII CHR8 PRO
                     /label=ZM-U6\POLIII\CHR8\PRO
                     /note="Maize U6 PolIII promoter from B73 chromosome 8."
                     /note="/vntifkey=30"
                     /db_xref="VRFEATURE:F000465"
                     /db_xref="VRROLE:promoter"
     misc_feature    1103..1122
                     /label=Cas9 R Promoter sgRNA Target 1
                     /label=Cas9 R Promoter Target 1
     misc_feature    1123..1199
                     /label=Cas9 sgRNA
                     /label=GUIDE\RNA
                     /note="Guide RNA (gRNA) referes to engineered 
                     crRNA-tracrRNA fusion transcripts used to direct Cas9 to 
                     cleave sequence of interest. Cas9, crRNA, and tracrRNA are 
                     components of the CRISPR system of Streptococcus pyogenes, 
                     Science 339:823-826."
                     /note="/vntifkey=21"
                     /db_xref="VRFEATURE:F000461"
                     /db_xref="VRROLE:sequence_feature"
     terminator      1200..1207
                     /label=ZM-U6 POLIII CHR8 TERM
                     /label=ZM-U6\POLIII\CHR8\TERM
                     /note="Maize U6 PolIII terminator from B73 chromosome 8."
                     /note="/vntifkey=43"
                     /db_xref="VRFEATURE:F000466"
                     /db_xref="VRROLE:terminator"
     promoter        complement(1662..1681)
                     /label=T3 PRO
                     /label=T3\PRO
                     /note="/vntifkey=30"
                     /db_xref="VRFEATURE:F004862"
                     /db_xref="VRROLE:promoter"
     rep_origin      complement(1918..2817)
                     /direction=LEFT
                     /label=COLE1 ORI
                     /label=COLE1\ORI
                     /note="/vntifkey=33"
                     /db_xref="VRFEATURE:F000006"
                     /db_xref="VRROLE:origin_of_replication"
     CDS             complement(2863..3723)
                     /codon_start=1
                     /label=AMP
                     /note="/vntifkey=4"
                     /db_xref="VRFEATURE:F000003"
                     /db_xref="VRROLE:CDS"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(3854..4309)
                     /direction=LEFT
                     /label=F1 ORI
                     /label=F1\ORI
                     /note="REPLICON; PARENT SOURCE=7770"
                     /note="/vntifkey=33"
                     /db_xref="VRFEATURE:F000456"
                     /db_xref="VRROLE:origin_of_replication"
ORIGIN
        1 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg
       61 ggtaccgggc cccccctcga ggtcgacggt atcgataagc tttgagagta caatgatgaa
      121 cctagattaa tcaatgccaa agtctgaaaa atgcaccctc agtctatgat ccagaaaatc
      181 aagattgctt gaggccctgt tcggttgttc cggattagag ccccggatta attcctagcc
      241 ggattacttc tctaatttat atagattttg atgagctgga atgaatcctg gcttattccg
      301 gtacaaccga acaggccctg aaggatacca gtaatcgctg agctaaattg gcatgctgtc
      361 agagtgtcag tattgcagca aggtagtgag ataaccggca tcatggtgcc agtttgatgg
      421 caccattagg gttagagatg gtggccatgg gcgcatgtcc tggccaactt tgtatgatat
      481 atggcagggt gaataggaaa gtaaaattgt attgtaaaaa gggatttctt ctgtttgtta
      541 gcgcatgtac aaggaatgca agttttgagc gagggggcat caaagatctg gctgtgtttc
      601 cagctgtttt tgttagcccc atcgaatcct tgacataatg atcccgctta aataagcaac
      661 ctcgcttgta tagttccttg tgctctaaca cacgatgatg ataagtcgta aaatagtggt
      721 gtccaaagaa tttccaggcc cagttgtaaa agctaaaatg ctattcgaat ttctactagc
      781 agtaagtcgt gtttagaaat tattttttta tatacctttt ttccttctat gtacagtagg
      841 acacagtgtc agcgccgcgt tgacggagaa tatttgcaaa aaagtaaaag agaaagtcat
      901 agcggcgtat gtgccaaaaa cttcgtcaca gagagggcca taagaaacat ggcccacggc
      961 ccaatacgaa gcaccgcgac gaagcccaaa cagcagtccg taggtggagc aaagcgctgg
     1021 gtaatacgca aacgttttgt cccaccttga ctaatcacaa gagtggagcg taccttataa
     1081 accgagccgc aagcaccgaa ttgagctcca ccaaagacaa aggttttaga gctagaaata
     1141 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     1201 tttttttgcg gccgcgaatt cctgcagggc cctcttgtcg gaccagttgc ccaccacgtt
     1261 ggtgagctcg gtgaggccct tcattgagag gaaggaggtc atgaggtgcc taccgatgtg
     1321 ggacttgggg ccgttcttga tggcgaagat ggagtagggg gcgttcttct tgagggcctt
     1381 gttgtaggac ctcacgaggt tgtccttgag gagctggtac tcctgcttgt tggaggagga
     1441 gttgccggtc ctgttcaccc tcttgagcac gggctctgag ttcctgagga actcgtcgag
     1501 gtacacgagg gggtcgatcc tgccgcgagc ggagaagaag tagatgtgcc tggacacgga
     1561 ggtcttggtc tcggtcacga ggcactggat gatcacgccg aggtacttgt tctgactagt
     1621 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     1681 ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1741 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1801 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1861 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1921 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1981 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2041 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2101 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2161 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2221 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2281 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2341 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2401 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2461 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2521 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2581 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2641 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2701 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2761 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2821 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2881 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2941 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3001 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3061 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3121 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3181 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3241 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3301 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3361 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3421 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3481 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3541 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3601 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3661 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3721 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3781 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3841 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3901 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3961 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4021 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4081 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4141 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4201 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4261 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     4321 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4381 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
//
