LOCUS       Exported                4440 bp ds-DNA     circular SYN 12-AUG-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    PV523
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        86..1085
                     /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_RNA        1086..1105
                     /label=Cas9 R Promoter sgRNA Target 2
                     /label=Cas9 R Promoter Target 2
                     /label=R Promoter Cas9 Target 2
                     /label=R-A7pl
                     /note="/vntifkey=53"
     misc_feature    1106..1182
                     /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      1183..1190
                     /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(1645..1664)
                     /label=T3 PRO
                     /label=T3\PRO
                     /note="/vntifkey=30"
                     /db_xref="VRFEATURE:F004862"
                     /db_xref="VRROLE:promoter"
     rep_origin      complement(1901..2800)
                     /direction=LEFT
                     /label=COLE1 ORI
                     /label=COLE1\ORI
                     /note="/vntifkey=33"
                     /db_xref="VRFEATURE:F000006"
                     /db_xref="VRROLE:origin_of_replication"
     CDS             complement(2846..3706)
                     /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(3837..4292)
                     /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 gacggccagt gaattgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct
       61 cgaggtcgac ggtatcgata agctttgaga gtacaatgat gaacctagat taatcaatgc
      121 caaagtctga aaaatgcacc ctcagtctat gatccagaaa atcaagattg cttgaggccc
      181 tgttcggttg ttccggatta gagccccgga ttaattccta gccggattac ttctctaatt
      241 tatatagatt ttgatgagct ggaatgaatc ctggcttatt ccggtacaac cgaacaggcc
      301 ctgaaggata ccagtaatcg ctgagctaaa ttggcatgct gtcagagtgt cagtattgca
      361 gcaaggtagt gagataaccg gcatcatggt gccagtttga tggcaccatt agggttagag
      421 atggtggcca tgggcgcatg tcctggccaa ctttgtatga tatatggcag ggtgaatagg
      481 aaagtaaaat tgtattgtaa aaagggattt cttctgtttg ttagcgcatg tacaaggaat
      541 gcaagttttg agcgaggggg catcaaagat ctggctgtgt ttccagctgt ttttgttagc
      601 cccatcgaat ccttgacata atgatcccgc ttaaataagc aacctcgctt gtatagttcc
      661 ttgtgctcta acacacgatg atgataagtc gtaaaatagt ggtgtccaaa gaatttccag
      721 gcccagttgt aaaagctaaa atgctattcg aatttctact agcagtaagt cgtgtttaga
      781 aattattttt ttatatacct tttttccttc tatgtacagt aggacacagt gtcagcgccg
      841 cgttgacgga gaatatttgc aaaaaagtaa aagagaaagt catagcggcg tatgtgccaa
      901 aaacttcgtc acagagaggg ccataagaaa catggcccac ggcccaatac gaagcaccgc
      961 gacgaagccc aaacagcagt ccgtaggtgg agcaaagcgc tgggtaatac gcaaacgttt
     1021 tgtcccacct tgactaatca caagagtgga gcgtacctta taaaccgagc cgcaagcacc
     1081 gaattgagtg ctataatatg agtgggtttt agagctagaa atagcaagtt aaaataaggc
     1141 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttttt gcggccgcga
     1201 attcctgcag ggccctcttg tcggaccagt tgcccaccac gttggtgagc tcggtgaggc
     1261 ccttcattga gaggaaggag gtcatgaggt gcctaccgat gtgggacttg gggccgttct
     1321 tgatggcgaa gatggagtag ggggcgttct tcttgagggc cttgttgtag gacctcacga
     1381 ggttgtcctt gaggagctgg tactcctgct tgttggagga ggagttgccg gtcctgttca
     1441 ccctcttgag cacgggctct gagttcctga ggaactcgtc gaggtacacg agggggtcga
     1501 tcctgccgcg agcggagaag aagtagatgt gcctggacac ggaggtcttg gtctcggtca
     1561 cgaggcactg gatgatcacg ccgaggtact tgttctgact agttctagag cggccgccac
     1621 cgcggtggag ctccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat
     1681 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1741 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1801 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1861 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1921 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1981 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2041 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2101 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2161 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2221 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2281 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2341 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2401 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2461 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2521 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2581 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2641 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2701 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2761 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2821 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2881 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2941 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3001 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3061 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3121 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3181 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3241 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3301 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3361 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3421 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3481 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3541 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3601 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3661 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3721 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3781 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     3841 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3901 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3961 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4021 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4081 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4141 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4201 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4261 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     4321 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     4381 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
//
