LOCUS       V015886                 5256 bp    DNA     circular SYN 01-FEB-2020
DEFINITION  Exported.
ACCESSION   V015886
VERSION     V015886
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5256)
  AUTHORS   Gunadi A, Orchard N, Qu F, Finer J.
  TITLE     Enhanced CRISPR/Cas9 mediated homology-independent targeted
            integration in plant cotyledonary tissues
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5256)
  AUTHORS   Gunadi A, Orchard N, Qu F, Finer J.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2019) Horticulture and Crop Science, The Ohio
            State University, 216 Williams Hall, 1680 Madison Ave., Wooster, OH
            44691, USA
REFERENCE   3  (bases 1 to 5256)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-JUN-2019) Horticulture and Crop Science, The Ohio State
            University, 216 Williams Hall, 1680 Madison Ave., Wooster, OH 44691,
            USA"
FEATURES             Location/Qualifiers
     source          1..5256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    32..53
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        68..98
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    106..122
                     /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     130..146
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        254..925
                     /label="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     regulatory      924..1067
                     /note="5' UTR enhancer sequence from Tobacco Etch Virus
                     (TEV)"
                     /regulatory_class="enhancer"
     CDS             1068..1568
                     /note="RNA silencing suppressor p19 from Tomato bushy stunt
                     virus (strain Cherry). Accession#: P11690"
                     /label="RNA silencing suppressor p19"
     regulatory      1572..1783
                     /label="CaMV35S terminator"
                     /note="CaMV35S terminator"
                     /regulatory_class="terminator"
     gene            1784..2476
                     /gene="hygR"
                     /label="hygR"
     CDS             1784..2476
                     /codon_start=1
                     /transl_table=11
                     /gene="hygR"
                     /product="HygR"
                     /label="hygR"
                     /protein_id="QHO64459.1"
                     /translation="MQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIAD
                     PHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVI
                     DWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQL
                     YQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRP
                     STRPRAKE"
     terminator      2523..2775
                     /label="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2784..2800)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3276..3380
                     /label="AmpR promoter"
     CDS             3381..4238
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4412..5000
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
       61 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      121 aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtca
      181 acatggtgga gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag
      241 accaaagggc aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc
      301 attgcccagc tatctgtcac tttattgtga agatagtgga aaaggaaggt ggctcctaca
      361 aatgccatca ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc
      421 ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt
      481 cttcaaagca agtggattga tgtgataaca tggtggagca cgacacactt gtctactcca
      541 aaaatatcaa agatacagtc tcagaagacc aaagggcaat tgagactttt caacaaaggg
      601 taatatccgg aaacctcctc ggattccatt gcccagctat ctgtcacttt attgtgaaga
      661 tagtggaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggccatcg
      721 ttgaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
      781 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca
      841 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
      901 gaagttcatt tcatttggag aggacctcga gaattctcaa cacaacatat acaaaacaaa
      961 cgaatctcaa gcaatcaagc attctacttc tattgcagca atttaaatca tttcttttaa
     1021 agcaaaagca attttctgaa aattttcacc atttacgaac gatagccatg gaacgagcta
     1081 tacaaggaaa cgacgctagg gaacaagcta acagtgaacg ttgggatgga ggatcaggag
     1141 gtaccacttc tcccttcaaa cttcctgacg aaagtccgag ttggactgag tggcggctac
     1201 ataacgatga gacgaattcg aatcaagata atccccttgg tttcaaggaa agctggggtt
     1261 tcgggaaagt tgtatttaag agatatctca gatacgacag gacggaagct tcactgcaca
     1321 gagtccttgg atcttggacg ggagattcgg ttaactatgc agcatctcga tttttcggtt
     1381 tcgaccagat cggatgtacc tatagtattc ggtttcgagg agttagtatc accgtttctg
     1441 gagggtcgcg aactcttcag catctctgtg agatggcaat tcggtctaag caagaactgc
     1501 tacagcttgc cccaatcgaa gtggaaagta atgtatcaag aggatgccct gaaggtactg
     1561 agaccttcta gagtccgcaa aaatcaccag tctctctcta caaatctatc tctctctatt
     1621 tttctccaga ataatgtgtg agtagttccc agataaggga attagggttc ttatagggtt
     1681 tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt
     1741 ctatcaataa aatttctaat tcctaaaacc aaaatccagt gacctgcagc cggtcgcgga
     1801 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     1861 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     1921 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     1981 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     2041 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     2101 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     2161 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     2221 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     2281 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     2341 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     2401 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     2461 gagggcaaag gaatagagta gatgccgacc gggatccccg ggtaccgagc tcgaatttcc
     2521 ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg
     2581 cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat
     2641 gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat
     2701 acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat
     2761 ctatgttact agatcgggaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2821 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2881 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2941 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     3001 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3061 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3121 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     3181 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     3241 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3301 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3361 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3421 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3481 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3541 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3601 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3661 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3721 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3781 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3841 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3901 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3961 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4021 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4081 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4141 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4201 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4261 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4321 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4381 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4441 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4501 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4561 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4621 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4681 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4741 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4801 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4861 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4921 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4981 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5041 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5101 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5161 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5221 cgattcatta atgcagctgg cacgacaggt ttcccg
//
