LOCUS       62056_1186        4238 bp DNA     circular SYN 01-FEB-2020
DEFINITION  Cloning vector HITI-GmScreaM8-recipient, complete sequence.
ACCESSION   MN043944
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4238)
  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  JOURNAL   Unpublished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 4238)
  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 4238)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      405..1978
                     /label=GmScreamM8 promoter from soybean
                     /note="GmScreamM8 promoter from soybean"
                     /regulatory_class="promoter"
     misc_feature    1979..2001
                     /note="directional target for CRISPR/Cas9-mediated
                     double-strand break"
     primer_bind     complement(2017..2033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2041..2057)
                     /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)."
     promoter        complement(2065..2095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2110..2131)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2419..3007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3181..4038)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4039..4143)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttttacgt gtgctgtgag
      421 acattatcat caattgtgtt gtatatgata tatagatata gatatataaa tatatagatt
      481 gagtgatata atatatttaa aatataaatt atatatatgt tttaatatat ttttgcatat
      541 atatatatat ttgtaaaaac tagaagtatt ttttcatgag ataattatta tcgagttgaa
      601 taagtctatt atttgtgaga gccaaccata tttatatatg tgattaaatt ttatctttgt
      661 gaaattaaaa ataataaaaa ataccttaaa atcataataa tagaaaaact tatatttata
      721 atttaccatt atacttaaaa aaaattaaat aaatattata aatataaata ctatcgagta
      781 atggccgcgc tagggttttt gagaaaaaat cttcccacgc actcaactgc actgtacggc
      841 gtcgttttca cagccgcata atagaagccg cgttccccaa cccttcctca caacattctc
      901 ggaccctcca gcaccgtcac ccaaacaaat atccacgcgg tagtaggcgc gtgaaacaaa
      961 ctctaatccg aactacgaga cgtgagaagc acgcgcttta gcgagcgttt caattgtcgc
     1021 tacgaaagca gagaaggata caaacggaac tagggtaaat tagtaagggt aatttcgtaa
     1081 acagaagaaa agagttgtag ctataaataa accctctaac cctcgtcgca ttacttctct
     1141 tcacaccttt gttcactctt cttctcttgc ggctagggtt ttagcgcagc ttcttctagg
     1201 ttcgttatct accaccgttc tatggatttt attccttcta ttcgtgttta ttctattggt
     1261 ttatgttgct tgcaatatgt tttttctgaa tctgtcgtcg ttgtcttcaa ttttatccat
     1321 gtttcagaga tcaattttgt ttgtgtagta tgtgcttatt cttcttcttt tcgttcgagt
     1381 tgttaataac ggtgctatgg tgttttcaaa agtgtttttt ttattacttt tgatttaaag
     1441 tttttttggt aaggctttta tttgcttgtt atattcaaat ctttggatcc agatcttata
     1501 taagtttttg gttcaagaaa gtttttggtt actgatgaat agatctatta actgttactt
     1561 taatcgattc aagctaaagt tttttggtta ctgatgaata gatctattat ctgttacttt
     1621 taatcggttc aagctcaagt tttttggtta ctgatgaata gatctatata cgtcacagtg
     1681 tgctaaacat gcccttgttt tatctcgatc ttatgtatgg gagtgccata aattttgtta
     1741 tgtctatttt tttatctgtt ggaatcatac tgagtttgat gcgttacgat tgagcatacc
     1801 tatttttggg cttgttgtat ggtgggtatt tagatcttaa tctttttatg cttatgaaag
     1861 gttttgtaat gacaaaggtc ttaatgttgt taaactttta tttttacttt atatggtgtg
     1921 ttgatgtgtt atggttttga caactttttt tttttctgga tttttgcaga tttaaggaat
     1981 ttagctgaag caaagcaatg ggaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     2041 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2101 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2161 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2221 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2281 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2341 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2401 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2461 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2521 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2581 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2641 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2701 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2761 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2821 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     2881 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2941 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3001 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3061 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3121 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3181 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3241 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3301 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3361 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3421 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3481 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3541 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3601 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3661 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3721 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3781 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3841 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3901 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3961 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4021 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4081 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4141 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4201 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
