LOCUS       62056_19680        9436 bp DNA     circular SYN 31-AUG-2021
DEFINITION  Cloning vector pSH179, complete sequence.
ACCESSION   MT670352
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9436)
  AUTHORS   Hiekel SSH., Budhagatapalli NNB.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2020) Physiology and Cell Biology/Plant 
            Reproductive Biology, Leibniz Institute of Plant Genetics and Crop 
            Plant Research (IPK), Corrensstrasse 3, Gatersleben, Saxony-Anhalt 
            06466, Germany
REFERENCE   2  (bases 1 to 9436)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (25-JUN-2020) Physiology and Cell Biology/Plant Reproductive 
            Biology, Leibniz Institute of Plant Genetics and Crop Plant Research
            (IPK), Corrensstrasse 3, Gatersleben, Saxony-Anhalt 06466, Germany"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..25)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     regulatory      126..488
                     /label=TaU6p
                     /note="TaU6p"
                     /regulatory_class="promoter"
     misc_RNA        510..585
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    586..592
                     /label=poly-T signal
                     /note="poly-T signal"
     promoter        710..1385
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     CDS             1561..5661
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     terminator      5754..6006
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6015..6031)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6170..6344
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    complement(6494..6518)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(6797..6824)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(6916..7002)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(7171..7606)
                     /direction=LEFT
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      complement(7721..8309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8402..9208)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagtttg taggccctta aggccatgaa
       61 actgaaggcg ggaaacgaca atctgatcca agctcaagct aagctcacgt gacggaatta
      121 agcttgacca agcccgttat tctgacagtt ctggtgctca acacatttat atttatcaag
      181 gagcacattg ttactcactg ctaggaggga atcgaactag gaatattgat cagaggaact
      241 acgagagagc tgaagataac tgccctctag ctctcactga tctgggtcgc atagtgagat
      301 gcagcccacg tgagttcagc aacggtctag cgctgggctt ttaggcccgc atgatcgggc
      361 ttttgtcggg tggtcgacgt gttcacgatt ggggagagca acgcagcagt tcctcttagt
      421 ttagtcccac ctcgcctgtc cagcagagtt ctgaccggtt tataaactcg cttgctgcat
      481 cagacttgcg agaccaacac aaggtctcgg ttttagagct agaaatagca agttaaaata
      541 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttaagcttcg
      601 aattcaagct ggagctcatg ttactagatc gggccaacat ggtggagcac gacactctcg
      661 tctactccaa gaatatcaaa gatacagtct cagaagacca aagggctatt gagacttttc
      721 aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc tgtcacttca
      781 tcaaaaggac agtagaaaag gaaggtggca cctacaaatg ccatcattgc gataaaggaa
      841 aggctatcgt tcaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga
      901 ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
      961 aacatggtgg agcacgacac tctcgtctac tccaagaata tcaaagatac agtctcagaa
     1021 gaccaaaggg ctattgagac ttttcaacaa agggtaatat cgggaaacct cctcggattc
     1081 cattgcccag ctatctgtca cttcatcaaa aggacagtag aaaaggaagg tggcacctac
     1141 aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctgc cgacagtggt
     1201 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg
     1261 tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc
     1321 cactatcctt cgcaagacct tcctctatat aaggaagttc atttcatttg gagaggacac
     1381 gctgaaatca ccagtctctc tctacaaatc tatctctctc gagcactgcg gccgctctag
     1441 atggattaca aggaccacga cggggattac aaggaccacg acattgatta caaggatgat
     1501 gatgacaaga tggctccgaa gaagaagagg aaggttggca tccacggggt gccagctgct
     1561 gacaagaagt actcgatcgg cctcgatatt gggactaact ctgttggctg ggccgtgatc
     1621 accgacgagt acaaggtgcc ctcaaagaag ttcaaggtcc tgggcaacac cgatcggcat
     1681 tccatcaaga agaatctcat tggcgctctc ctgttcgaca gcggcgagac ggctgaggct
     1741 acgcggctca agcgcaccgc ccgcaggcgg tacacgcgca ggaagaatcg catctgctac
     1801 ctgcaggaga ttttctccaa cgagatggcg aaggttgacg attctttctt ccacaggctg
     1861 gaggagtcat tcctcgtgga ggaggataag aagcacgagc ggcatccaat cttcggcaac
     1921 attgtcgacg aggttgccta ccacgagaag taccctacga tctaccatct gcggaagaag
     1981 ctcgtggact ccacagataa ggcggacctc cgcctgatct acctcgctct ggcccacatg
     2041 attaagttca ggggccattt cctgatcgag ggggatctca acccggacaa tagcgatgtt
     2101 gacaagctgt tcatccagct cgtgcagacg tacaaccagc tcttcgagga gaaccccatt
     2161 aatgcgtcag gcgtcgacgc gaaggctatc ctgtccgcta ggctctcgaa gtctcggcgc
     2221 ctcgagaacc tgatcgccca gctgccgggc gagaagaaga acggcctgtt cgggaatctc
     2281 attgcgctca gcctggggct cacgcccaac ttcaagtcga atttcgatct cgctgaggac
     2341 gccaagctgc agctctccaa ggacacatac gacgatgacc tggataacct cctggcccag
     2401 atcggcgatc agtacgcgga cctgttcctc gctgccaaga atctgtcgga cgccatcctc
     2461 ctgtctgata ttctcagggt gaacaccgag attacgaagg ctccgctctc agcctccatg
     2521 atcaagcgct acgacgagca ccatcaggat ctgaccctcc tgaaggcgct ggtcaggcag
     2581 cagctccccg agaagtacaa ggagatcttc ttcgatcagt cgaagaacgg ctacgctggg
     2641 tacattgacg gcggggcctc tcaggaggag ttctacaagt tcatcaagcc gattctggag
     2701 aagatggacg gcacggagga gctgctggtg aagctcaatc gcgaggacct cctgaggaag
     2761 cagcggacat tcgataacgg cagcatccca caccagattc atctcgggga gctgcacgct
     2821 atcctgagga ggcaggagga cttctaccct ttcctcaagg ataaccgcga gaagatcgag
     2881 aagattctga ctttcaggat cccgtactac gtcggcccac tcgctagggg caactcccgc
     2941 ttcgcttgga tgacccgcaa gtcagaggag acgatcacgc cgtggaactt cgaggaggtg
     3001 gtcgacaagg gcgctagcgc tcagtcgttc atcgagagga tgacgaattt cgacaagaac
     3061 ctgccaaatg agaaggtgct ccctaagcac tcgctcctgt acgagtactt cacagtctac
     3121 aacgagctga ctaaggtgaa gtatgtgacc gagggcatga ggaagccggc tttcctgtct
     3181 ggggagcaga agaaggccat cgtggacctc ctgttcaaga ccaaccggaa ggtcacggtt
     3241 aagcagctca aggaggacta cttcaagaag attgagtgct tcgattcggt cgagatctct
     3301 ggcgttgagg accgcttcaa cgcctccctg gggacctacc acgatctcct gaagatcatt
     3361 aaggataagg acttcctgga caacgaggag aatgaggata tcctcgagga cattgtgctg
     3421 acactcactc tgttcgagga ccgggagatg atcgaggagc gcctgaagac ttacgcccat
     3481 ctcttcgatg acaaggtcat gaagcagctc aagaggagga ggtacaccgg ctgggggagg
     3541 ctgagcagga agctcatcaa cggcattcgg gacaagcagt ccgggaagac gatcctcgac
     3601 ttcctgaaga gcgatggctt cgcgaaccgc aatttcatgc agctgattca cgatgacagc
     3661 ctcacattca aggaggatat ccagaaggct caggtgagcg gccaggggga ctcgctgcac
     3721 gagcatatcg cgaacctcgc tggctcgcca gctatcaaga aggggattct gcagaccgtg
     3781 aaggttgtgg acgagctggt gaaggtcatg ggcaggcaca agcctgagaa catcgtcatt
     3841 gagatggccc gggagaatca gaccacgcag aagggccaga agaactcacg cgagaggatg
     3901 aagaggatcg aggagggcat taaggagctg gggtcccaga tcctcaagga gcacccggtg
     3961 gagaacacgc agctgcagaa tgagaagctc tacctgtact acctccagaa tggccgcgat
     4021 atgtatgtgg accaggagct ggatattaac aggctcagcg attacgacgt cgatcatatc
     4081 gttccacagt cattcctgaa ggatgactcc attgacaaca aggtcctcac caggtcggac
     4141 aagaaccggg gcaagtctga taatgttcct tcagaggagg tcgttaagaa gatgaagaac
     4201 tactggcgcc agctcctgaa tgccaagctg atcacgcagc ggaagttcga taacctcaca
     4261 aaggctgaga ggggcgggct ctctgagctg gacaaggcgg gcttcatcaa gaggcagctg
     4321 gtcgagacac ggcagatcac taagcacgtt gcgcagattc tcgactcacg gatgaacact
     4381 aagtacgatg agaatgacaa gctgatccgc gaggtgaagg tcatcaccct gaagtcaaag
     4441 ctcgtctccg acttcaggaa ggatttccag ttctacaagg ttcgggagat caacaattac
     4501 caccatgccc atgacgcgta cctgaacgcg gtggtcggca cagctctgat caagaagtac
     4561 ccaaagctcg agagcgagtt cgtgtacggg gactacaagg tttacgatgt gaggaagatg
     4621 atcgccaagt cggagcagga gattggcaag gctaccgcca agtacttctt ctactctaac
     4681 attatgaatt tcttcaagac agagatcact ctggccaatg gcgagatccg gaagcgcccc
     4741 ctcatcgaga cgaacggcga gacgggggag atcgtgtggg acaagggcag ggatttcgcg
     4801 accgtcagga aggttctctc catgccacaa gtgaatatcg tcaagaagac agaggtccag
     4861 actggcgggt tctctaagga gtcaattctg cctaagcgga acagcgacaa gctcatcgcc
     4921 cgcaagaagg actgggatcc gaagaagtac ggcgggttcg acagccccac tgtggcctac
     4981 tcggtcctgg ttgtggcgaa ggttgagaag ggcaagtcca agaagctcaa gagcgtgaag
     5041 gagctgctgg ggatcacgat tatggagcgc tccagcttcg agaagaaccc gatcgatttc
     5101 ctggaggcga agggctacaa ggaggtgaag aaggacctga tcattaagct ccccaagtac
     5161 tcactcttcg agctggagaa cggcaggaag cggatgctgg cttccgctgg cgagctgcag
     5221 aaggggaacg agctggctct gccgtccaag tatgtgaact tcctctacct ggcctcccac
     5281 tacgagaagc tcaagggcag ccccgaggac aacgagcaga agcagctgtt cgtcgagcag
     5341 cacaagcatt acctcgacga gatcattgag cagatttccg agttctccaa gcgcgtgatc
     5401 ctggccgacg cgaatctgga taaggtcctc tccgcgtaca acaagcaccg cgacaagcca
     5461 atcagggagc aggctgagaa tatcattcat ctcttcaccc tgacgaacct cggcgcccct
     5521 gctgctttca agtacttcga cacaactatc gatcgcaaga ggtacacaag cactaaggag
     5581 gtcctggacg cgaccctcat ccaccagtcg attaccggcc tctacgagac gcgcatcgac
     5641 ctgtctcagc tcgggggcga caagcggcca gcggcgacga agaaggcggg gcaggcgaag
     5701 aagaagaagt gagctcaggc ctccatacaa gtattgggga tccgaatttc cccgatcgtt
     5761 caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta
     5821 tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt
     5881 tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag
     5941 aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac
     6001 tagatcggga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     6061 acccaactta atcgccttgc agcacatccc cctttcgcca ggggtaccag gccgccatgg
     6121 ccaggtaccg ccccgtccgg tcctgcccgt caccgagatt tgactcgagt ttctccataa
     6181 taatgtgtga gtagttccca gataagggaa ttagggttcc tatagggttt cgctcatgtg
     6241 ttgagcatat aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa
     6301 atttctaatt cctaaaacca aaatccagta ctaaaatcca gatcccccga attaattcgg
     6361 cgttaattca gtacattaaa aacgtccgca atgtgttatt aagttgtcac tagtcaggtt
     6421 aactcaattc ggcgttaatt cagtacatta aaaacgtccg caatgtgtta ttaagttgtc
     6481 taagcgtcaa tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg
     6541 accggcagct cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg
     6601 tcagcgggag agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa
     6661 gaacggcaac taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg
     6721 attgtaacga tgacagagcg ttgctgcctg tgatcactta agtaactaac taacaggaag
     6781 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     6841 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     6901 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     6961 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     7021 ctctcgctta gtagttagac gtccccgaga tccatgctag accatgaatc cagaagcccg
     7081 agaggttgcc gcctttcggg ctttttcttt ttcaaaaaaa aaaatttata aaacgatctg
     7141 ttgcggccgg ccgccgggtt gtgggcaaag gcgctcgacg gtgggcaacc gcttgcggtt
     7201 gtccacgggc ggagccggtg cgcgtagcgc attgtccaca agccaagggc gaccaataat
     7261 tgatatatat attcataatt gaaaagctaa ttgaacatac tacttgctgt aactacttgc
     7321 cggagcgagg ggtgtttgca agctgttgat ctgaaagggc tattagcgtt ctcacgtgcc
     7381 tttttgatta gcgatttcac gtgaccttat tagcgatttc acgtactccg attagcgatt
     7441 tcacgtaccc tgattagcga tttcacgtgg atagtttttg gagcgggccg gaaagccccg
     7501 tgaatcaagg ctttgcgggg cattagcggt ttcacgtgga taactaccct ctatccacag
     7561 gcttccgggg ataaaaaagc ccgctcgacg gcgggctgtt ggatggggat ctagcggtaa
     7621 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7681 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7741 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7801 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7861 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7921 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7981 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     8041 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     8101 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8161 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8221 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8281 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct accgggtctg
     8341 acgctcagtg gaacggggcc caatctgaat aatgttacaa ccaattaacc aattctgatt
     8401 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     8461 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     8521 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     8581 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     8641 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     8701 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     8761 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     8821 gcaaccggcg cagggacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     8881 cttctaatac ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat
     8941 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     9001 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     9061 actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat
     9121 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     9181 tcgacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     9241 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     9301 gagacacggg ccagagctgc agtttgatcc cgaggggaac cctgtggttg acatgcacat
     9361 acaaatggac gaacggataa accttttcac gcccttttaa atatccgtta ttctaataaa
     9421 cgctcttttc tcttag
//
