LOCUS       62056_66        9396 bp DNA     circular SYN 19-MAR-2020
DEFINITION  Cloning vector 1-UBI_Cas9_PinII, complete sequence.
ACCESSION   MN294713
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9396)
  AUTHORS   Gao H.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-AUG-2019) Molecular Engineering in AST, Corteva 
            Agriscience, 8305 NW 62nd Ave., Johnston, IA 50131, USA
REFERENCE   2  (bases 1 to 9396)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (08-AUG-2019) Molecular Engineering in AST, Corteva Agriscience, 
            8305 NW 62nd Ave., Johnston, IA 50131, USA"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Vector NTI v. 10
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      13..322
                     /regulatory_class="terminator"
     protein_bind    complement(378..477)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(495..513)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(518..534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             647..1453
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1603..2191
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2521..2548)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(2640..2726)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     2790..2806
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2822..2921
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3052..5046
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             5073..5093
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     intron          5472..5660
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             6891..9389
                     /label=Cas9(C)
                     /note="C-terminal portion of Streptococcus pyogenes Cas9 
                     (Zetsche et al., 2015)"
ORIGIN
        1 cacatggtta acctagactt gtccatcttc tggattggcc aacttaatta atgtatgaaa
       61 taaaaggatg cacacatagt gacatgctaa tcactataat gtgggcatca aagttgtgtg
      121 ttatgtgtaa ttactagtta tctgaataaa agagaaagag atcatccata tttcttatcc
      181 taaatgaatg tcacgtgtct ttataattct ttgatgaacc agatgcattt cattaaccaa
      241 atccatatac atataaatat taatcatata taattaatat caattgggtt agcaaaacaa
      301 atctagtcta ggtgtgtttt gcgaattgcg gccagcttgg tcacccggtc cgggcctaga
      361 aggccgatct cccgggcacc cagctttctt gtacaaagtg ggcattataa gaaagcattg
      421 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
      481 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
      541 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
      601 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
      661 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
      721 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
      781 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
      841 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
      901 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
      961 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     1021 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     1081 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     1141 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     1201 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     1261 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     1321 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     1381 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     1441 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     1501 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     1561 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1621 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1681 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1741 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1801 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1861 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1921 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1981 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2041 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2101 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2161 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2221 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2281 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2341 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     2401 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     2461 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     2521 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     2581 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     2641 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     2701 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     2761 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
     2821 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
     2881 atgctttttt ataatgccaa gtttgtacaa aaaagcaggc tccggccaga atggcccgga
     2941 ccgggttacc gaattcgcct agaaggccat ttaaatcctg aggatctggt cttcctaagg
     3001 acccgggata tcggaccgat taaactttaa ttcggtccga agcttgcatg cctgcagtgc
     3061 agcgtgaccc ggtcgtgccc ctctctagag ataatgagca ttgcatgtct aagttataaa
     3121 aaattaccac atattttttt tgtcacactt gtttgaagtg cagtttatct atctttatac
     3181 atatatttaa actttactct acgaataata taatctatag tactacaata atatcagtgt
     3241 tttagagaat catataaatg aacagttaga catggtctaa aggacaattg agtattttga
     3301 caacaggact ctacagtttt atctttttag tgtgcatgtg ttctcctttt tttttgcaaa
     3361 tagcttcacc tatataatac ttcatccatt ttattagtac atccatttag ggtttagggt
     3421 taatggtttt tatagactaa tttttttagt acatctattt tattctattt tagcctctaa
     3481 attaagaaaa ctaaaactct attttagttt ttttatttaa taatttagat ataaaataga
     3541 ataaaataaa gtgactaaaa attaaacaaa taccctttaa gaaattaaaa aaactaagga
     3601 aacatttttc ttgtttcgag tagataatgc cagcctgtta aacgccgtcg acgagtctaa
     3661 cggacaccaa ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag acggcacggc
     3721 atctctgtcg ctgcctctgg acccctctcg agagttccgc tccaccgttg gacttgctcc
     3781 gctgtcggca tccagaaatt gcgtggcgga gcggcagacg tgagccggca cggcaggcgg
     3841 cctcctcctc ctctcacggc acggcagcta cgggggattc ctttcccacc gctccttcgc
     3901 tttcccttcc tcgcccgccg taataaatag acaccccctc cacaccctct ttccccaacc
     3961 tcgtgttgtt cggagcgcac acacacacaa ccagatctcc cccaaatcca cccgtcggca
     4021 cctccgcttc aaggtacgcc gctcgtcctc cccccccccc ctctctacct tctctagatc
     4081 ggcgttccgg tccatgcatg gttagggccc ggtagttcta cttctgttca tgtttgtgtt
     4141 agatccgtgt ttgtgttaga tccgtgctgc tagcgttcgt acacggatgc gacctgtacg
     4201 tcagacacgt tctgattgct aacttgccag tgtttctctt tggggaatcc tgggatggct
     4261 ctagccgttc cgcagacggg atcgatttca tgattttttt tgtttcgttg catagggttt
     4321 ggtttgccct tttcctttat ttcaatatat gccgtgcact tgtttgtcgg gtcatctttt
     4381 catgcttttt tttgtcttgg ttgtgatgat gtggtctggt tgggcggtcg ttctagatcg
     4441 gagtagaatt ctgtttcaaa ctacctggtg gatttattaa ttttggatct gtatgtgtgt
     4501 gccatacata ttcatagtta cgaattgaag atgatggatg gaaatatcga tctaggatag
     4561 gtatacatgt tgatgcgggt tttactgatg catatacaga gatgcttttt gttcgcttgg
     4621 ttgtgatgat gtggtgtggt tgggcggtcg ttcattcgtt ctagatcgga gtagaatact
     4681 gtttcaaact acctggtgta tttattaatt ttggaactgt atgtgtgtgt catacatctt
     4741 catagttacg agtttaagat ggatggaaat atcgatctag gataggtata catgttgatg
     4801 tgggttttac tgatgcatat acatgatggc atatgcagca tctattcata tgctctaacc
     4861 ttgagtacct atctattata ataaacaagt atgttttata attattttga tcttgatata
     4921 cttggatgat ggcatatgca gcagctatat gtggattttt ttagccctgc cttcatacgc
     4981 tatttatttg cttggtactg tttcttttgt cgatgctcac cctgttgttt ggtgttactt
     5041 ctgcaggtcg actctagagg atccccatgg ccccgaagaa gaagaggaag gtgcacatgg
     5101 ataagaagta cagcatcggc ctcgacatcg ggaccaacag cgtcggctgg gccgtcatca
     5161 ccgacgaata taaggtgccc agcaagaagt tcaaggtgct cgggaataca gaccgccaca
     5221 gcatcaagaa gaacctgatc ggcgccctcc tgttcgactc gggcgagacc gctgaggcca
     5281 ccagactaaa gaggaccgct cgccgccgct acacccgccg caagaaccgc atatgctacc
     5341 tccaggagat cttcagcaac gagatggcca aggtggacga cagcttcttc caccgccttg
     5401 aggagtcgtt cctcgtggag gaggacaaga agcatgagag gcacccgatc ttcgggaaca
     5461 tcgtggacga ggtaagtttc tgcttctacc tttgatatat atataataat tatcattaat
     5521 tagtagtaat ataatatttc aaatattttt ttcaaaataa aagaatgtag tatatagcaa
     5581 ttgcttttct gtagtttata agtgtgtata ttttaattta taacttttct aatatatgac
     5641 caaaacatgg tgatgtgcag gtggcgtacc acgagaagta cccgacgatc taccacctcc
     5701 gcaagaagct ggtcgactcc acagacaagg ccgacctcag actgatctac ctggccctcg
     5761 cgcacatgat caagttccgc gggcacttcc tcatcgaggg cgacctgaac ccggacaact
     5821 ccgacgtcga caagctcttc atccagctgg tccagaccta caatcaactg ttcgaggaga
     5881 acccgatcaa cgcgtccggc gtggacgcga aggccatcct cagcgcgagg ctcagcaaat
     5941 caagacggct ggagaacctg atcgcccagc tcccaggcga gaagaaaaac ggcttgttcg
     6001 gcaacctgat cgcgctctcg ctcggcctca cgcccaactt caaatcaaac ttcgacctgg
     6061 ccgaggacgc gaaactgcag ctgtccaagg acacttacga cgacgacctc gacaacctgc
     6121 tggcgcaaat cggtgaccag tacgcagacc tcttcctggc cgccaagaac ctctcggacg
     6181 ccatcctgct gtccgatatc ctgagagtga atacggagat caccaaggcg ccgctcagcg
     6241 cctccatgat taaaaggtac gacgagcacc accaggacct gacgctgctc aaggccctgg
     6301 tgcgccagca gctccccgag aagtacaagg agatcttctt cgaccaatca aaaaacggct
     6361 acgccggcta catcgacggg ggcgcctccc aggaggagtt ctacaagttc atcaaaccaa
     6421 ttctcgagaa gatggacggc acggaggagc ttctcgtgaa gctcaaccgg gaggacctcc
     6481 tgaggaagca gaggacgttc gacaacggct cgataccgca tcagatccac ctgggcgagc
     6541 tccacgccat cctgcgccgg caggaggatt tctatccgtt cctcaaggac aacagggaga
     6601 agatcgagaa aattctgacg ttccgcatcc cgtactacgt gggccctctc gcgcgcggga
     6661 acagccggtt cgcctggatg actcggaagt cggaggagac gatcacgccg tggaacttcg
     6721 aggaggtggt ggacaagggc gcctccgccc agtcgttcat cgagcgcatg acgaacttcg
     6781 ataaaaatct gcccaatgaa aaagtgctcc cgaagcacag cctcctctac gagtacttca
     6841 cggtgtacaa cgagctcacg aaggtgaagt acgtgaccga gggtatgcgg aagccggcgt
     6901 tcctgagcgg cgagcagaag aaggccatcg tggacctcct cttcaagacg aaccggaaag
     6961 tcaccgtgaa gcaattaaag gaggactact tcaagaaaat agagtgcttc gacagcgtcg
     7021 agatctcggg cgtcgaggac aggttcaacg cgtcgctggg cacataccac gacctcctca
     7081 agatcattaa agacaaggac ttcctggaca acgaggagaa cgaggacatc ctcgaggaca
     7141 tcgtgctgac cctcaccctg tttgaggacc gggagatgat cgaggagcgc ctcaagacgt
     7201 acgctcacct tttcgacgac aaggtgatga aacagctgaa gcggcgccgc tacaccggat
     7261 ggggccggct ctcccgcaag ctcattaatg ggatcaggga caagcagtcc ggcaagacca
     7321 tactcgattt cctgaagagc gacggcttcg ccaaccggaa cttcatgcag ctcatccacg
     7381 acgactccct cactttcaag gaggacatcc agaaggccca ggtcagcgga cagggcgact
     7441 cgctccacga acacatcgcc aacctggccg ggtcgcctgc gattaaaaag ggaatccttc
     7501 agaccgtcaa ggtcgtggac gagctggtga aggtgatggg caggcacaag cccgaaaata
     7561 tcgtcattga gatggcccgg gagaaccaga ccacgcagaa aggccagaag aacagccggg
     7621 agcgcatgaa acggatcgag gagggtatca aggagctggg ctcgcagatc ctcaaggagc
     7681 accctgtgga aaatacccag ctgcagaatg aaaagctcta cctctactac ctccagaacg
     7741 gccgcgacat gtacgtggac caggagctgg acattaatcg cctctcggac tacgacgtcg
     7801 accacatcgt cccgcagtcc ttcctgaagg acgacagcat cgacaacaag gtcttgaccc
     7861 gctccgataa aaatcgcggg aagtccgaca acgtgccgtc ggaggaggtg gtcaagaaga
     7921 tgaaaaacta ctggcgccag ctgctcaacg ccaagctaat cacgcagcgc aagttcgaca
     7981 acctcaccaa ggccgaacgc ggcggtctct ccgagcttga taaggctggg ttcatcaaga
     8041 gacagctggt ggagacccgg cagatcacca agcatgtcgc ccagatcctg gactcgcgca
     8101 tgaatactaa gtacgatgaa aacgacaagc tcatccgcga ggtgaaggtg atcaccctga
     8161 agagcaagct ggtctcggac ttccggaagg acttccagtt ctacaaggtc cgggagatca
     8221 acaactacca ccacgcgcac gacgcctacc tgaacgcggt ggtgggcaca gcccttataa
     8281 agaagtaccc taagctcgag tccgagttcg tgtacggcga ctacaaggtg tacgacgtcc
     8341 gcaagatgat cgcgaagagc gagcaggaga tcgggaaggc caccgcaaaa tacttcttct
     8401 actccaacat catgaacttc ttcaagaccg agatcaccct ggccaacggg gagatccgca
     8461 agcgcccgct gattgagacg aacggagaga caggcgagat agtctgggac aagggcaggg
     8521 acttcgccac cgtgcgcaag gttctgtcca tgccgcaggt gaacatcgtg aagaagactg
     8581 aggtgcagac aggcggcttc tcgaaggagt ccatcctgcc caagcggaac agcgacaagc
     8641 tcatcgcgcg gaagaaggac tgggacccta aaaaatatgg cgggttcgac tcgcccaccg
     8701 tggcttactc ggtcctcgtg gtggccaagg tcgagaaggg caaaagcaag aagctgaaga
     8761 gcgtcaagga gctcctcggc atcaccatca tggagcggtc cagcttcgag aagaacccga
     8821 tcgacttcct cgaggcgaag ggatataagg aggtgaagaa ggacctcatc attaaactgc
     8881 cgaagtactc gctattcgaa ctggagaatg gtcgcaagag gatgctcgcg agcgctggcg
     8941 agctgcagaa agggaacgag ctggctctcc cgagcaagta cgtcaacttc ctctacctgg
     9001 cctcccacta tgaaaagctc aagggctcgc cggaggacaa cgagcagaag cagctgttcg
     9061 tcgagcagca caagcattac ctcgacgaga tcatcgagca gatctcggag ttcagcaagc
     9121 gcgtgatcct ggccgacgcc aacctcgaca aggtgctgtc cgcatataac aagcaccgcg
     9181 acaaaccaat acgggagcag gccgaaaata tcatccacct gttcaccctc acgaacctgg
     9241 gcgcccccgc cgcgttcaag tacttcgaca caaccatcga ccgcaagcgg tacacgagca
     9301 cgaaggaggt gctggacgcc acgttgattc accagtccat cacgggcctg tatgaaacaa
     9361 ggatcgatct cagccagctc ggcggcgact aggtac
//
