LOCUS       62056_10855        5622 bp DNA     circular SYN 01-FEB-2020
DEFINITION  Cloning vector pFLGA-GmubiXL-RGR, complete sequence.
ACCESSION   MN043940
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5622)
  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  JOURNAL   Unpublished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 5622)
  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 5622)
  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            "Unpublished"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..5622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..20)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        494..598
                     /label=AmpR promoter
     CDS             599..1456
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1630..2218
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2506..2527
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2542..2572
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2580..2596
                     /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     2604..2620
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2701..5125
                     /label=GmubiXL promoter from soybean
                     /note="GmubiXL promoter from soybean"
                     /regulatory_class="promoter"
     ncRNA           5140..5176
                     /label=hammerhead ribozyme
     misc_RNA        5197..5272
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           5273..5340
                     /label=HDV ribozyme
     terminator      5341..5593
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
       61 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
      121 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
      181 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
      241 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
      301 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
      361 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
      421 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggttctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     2461 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     2521 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     2581 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttat
     2641 taccctgtta tccctagata tcgattttgg ctaccttaac tctcttaagg tagcccatgg
     2701 acataagtct atctaaaatg gaaagattgt tacctaatct caaatcatat ttttcccatc
     2761 ataaaaaaaa atctcaaatc atgtgatctt gtcgatcatc atttaataca ttattaattc
     2821 ttctttttaa ttgaaaaatg tgattatatg ttatgtggag attagttcag accatgatcc
     2881 ttgaatatgt agtaagtttt tgtaatgttg agtttgagtt ctttgacact aggtccatgt
     2941 ttcccaaaat taagtgtaaa aaataaataa agaattaaaa gttgtgaaaa aaattgtcct
     3001 cgtttatttt tattattcag aattaatctt tattgagact aaaattattt acagaaagtt
     3061 aatacagcac taaaagttat tttatttttt tgagaaacaa aaactgaaat ctaaaaaaaa
     3121 ataagaaacc ggaaatctaa ttaacaaaaa tatatacggg ctaacatggt tatgggctta
     3181 ggctgcctca ttacccatta catgctctgt tcctaaaaaa aaaaagaagt ttgggccttt
     3241 atcctccagt aagtttttta attgggctca agaactttag gatttaggtg aggatcaatg
     3301 aaagtttata tattaggctt gatttttttt tcaatttacg taaaaatgaa agaaaaagtc
     3361 acactctata aactttattt ttttcaatat taatacattg ttattgtgga tatatcatta
     3421 gcggaaaagt aattctaata ccagatttaa ttgactttga ctaatttttt tagcggaata
     3481 aaagttttgn tactttgncg cgtgattttt aatnnaactt tggggataat ttcccctcca
     3541 attcagccca aaaaaaaaaa aactgannnc atatnttatt agtaggcaac ttgttcgtaa
     3601 atggtgtggc ttatccgaat ggaaatngga ttcgttttct ttaacatcat tattgttttt
     3661 gtcaatgagc tatcttttag tcttatgtta ttggtgaatc tgtccttaag ttgcatcatt
     3721 taacacatct cctcattaga gaaaaaaatt cttccctaaa cgattggtag taaaaacatc
     3781 taataagaaa taagaaagaa aaattaggaa aaaggaaagt tcattaaaaa aaatattttg
     3841 aattattttt taaaaaatat ctaaatattt tttaaatgaa taattttata taaactgtaa
     3901 ctaaatgtat acaagtaatg tatgttaaaa aaatacttga aaaatctact gaaaatatat
     3961 cttacaaggt gaaattaaat aagaaagaat ttagtggaat aattatgatt ttatttaaaa
     4021 aataattatt aaagattttt ttgctccata ataagaaaac ttttcaatta ttcttttctg
     4081 gtccataata aaaaaaatct agcatgacag cttttccata gatttttaat aatgtaaaag
     4141 cagccgactt caggcaatgg atagtggggc ccgtatcaac ttcggacgct ccacttgcaa
     4201 cggggtgggc ccaatataac aacgacgtcg taacagataa agcgaagctt gaaggtgcat
     4261 gtgactccgt caagattacg aaaccgccaa ctaccacgca aattgcaatt ctcaatttcc
     4321 tagaaggact ctccgaaaat gcatccaata ccaaatatta cccgtgtcat aggcaccaag
     4381 tgacaccata catgaacacg cgtcacaata tgactggaga agggttccac accttatgct
     4441 ataaaacgcc ccacacccct cctccttcct tcgcagttca attccaatat attccattct
     4501 ctctgtgtat ttccctacct ctcccttcaa ggttagtcga tttcttctgt ttttcttctt
     4561 cgttctttcc atgaattgtg tatgttcttt gatcaatacg atgttgattt gattgtgttt
     4621 tgtttggttt catcgatctt caattttcat aatcagattc agcttttatt atctttacaa
     4681 caacgtcctt aatttgatga ttctttaatc gtagatttgc tctaattaga gctttttcat
     4741 gtcagatccc tttacaacaa gccttaattg ttgattcatt aatcgtagat tagggctttt
     4801 ttcattgatt acttcagatc cgttaaacgt aaccatagat cagggctttt tcatgaatta
     4861 cttcagatcc gttaaacaac agccttattt tttatacttc tgtggttttt caagaaattg
     4921 ttcagatccg ttgacaaaaa gccttattcg ttgattctat atcgtttttc gagagatatt
     4981 gctcagatct gttagcaact gccttgtttg ttgattctat tgccgtggat tagggttttt
     5041 tttcacgaga ttgcttcaga tccgtactta agattacgta atggattttg attctgattt
     5101 atctgtgatt gttgactcga caggagcggc cgcctaaatc tgatgagtcc gtgaggacga
     5161 aacgagtaag ctcgtcattt agctgaagca aagcaagttt tagagctaga aatagcaagt
     5221 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcggccggca
     5281 tggtcccagc ctcctcgctg gcgccggctg ggcaacatgc ttcggcatgg cgaatgggac
     5341 gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg
     5401 atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc
     5461 atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac
     5521 gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct
     5581 atgttactag atccgatgat aagctgtcaa acatgagaat tc
//
