LOCUS       62056_1191        5290 bp DNA     circular SYN 01-FEB-2020
DEFINITION  Cloning vector HITI-GmScreamM8-reconstituted-GFPNOS, complete 
            sequence.
ACCESSION   MN043943
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5290)
  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  JOURNAL   Unpublished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 5290)
  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 5290)
  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            "Unpublished"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..5290
                     /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..2013
                     /label=5' HITI-1 junction
                     /note="5' HITI-1 junction"
     CDS             2014..2730
                     /label=EGFP
                     /note="enhanced GFP"
     terminator      2766..3018
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3048..3059
                     /label=3' HITI-1 junction
                     /note="3' HITI-1 junction"
     primer_bind     complement(3069..3085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3093..3109)
                     /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(3117..3147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3162..3183)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3471..4059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4233..5090)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5091..5195)
                     /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 tttaaggaaa
     1981 tttagctgaa gcaaagcttt gcttcagcta aatatggtga gcaagggcga ggagctgttc
     2041 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2101 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2161 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcac ctacggcgtg
     2221 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2281 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2341 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2401 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2461 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2521 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2581 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     2641 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2701 atcactcacg gcatggacga gctgtacaag taaagcggcc gcccggctgc actgaggcct
     2761 cagcagatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc
     2821 ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt
     2881 aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt
     2941 aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt
     3001 catctatgtt actagatccg atgataagct gtcaaacatg agaattccca ttgcaatggg
     3061 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3121 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3181 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3241 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3301 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3361 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3421 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3481 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3541 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3601 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3661 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3721 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3781 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3841 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3901 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3961 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4021 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4081 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4141 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4201 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4261 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4321 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4381 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4441 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4501 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4561 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4621 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4681 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4741 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4801 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4861 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4921 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4981 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5041 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5101 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5161 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5221 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5281 ccctttcgtc
//
