LOCUS       pACYC-T7-SpCas9-T7-EGFPgRNA1-T7term_(BPK848)        8020 bp    DNA     circular UNA 05-JAN-2022
DEFINITION  pACYCDUET-1.
ACCESSION   urn.local...jj-eb0j5d4
VERSION     urn.local...jj-eb0j5d4
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     protein_bind    5..25
                     /standard_name="lacO"
     misc_feature    58..63
                     /note="Geneious type: RBS"
                     /standard_name="RBS-1"
     CDS             71..4174
                     /standard_name="SpCas9"
     CDS             4175..4180
                     /standard_name="linker"
     CDS             4181..4201
                     /standard_name="NLS (SV40)"
     CDS             4202..4207
                     /standard_name="linker"
     CDS             4208..4273
                     /standard_name="3xFLAG"
     misc_feature    4277
                     /note="Geneious type: Editing History Insertion"
                     /standard_name
     misc_feature    4318..4337
                     /note="Geneious type: primer"
                     /standard_name="DuetUP2"
     misc_feature    complement(4318..4337)
                     /note="Geneious type: primer"
                     /standard_name="DuetDOWN1"
     regulatory      4343..4359
                     /note="Geneious type: promoter"
                     /standard_name="T7 promoter"
     misc_binding    4362..4381
     misc_RNA        4382..4458
                     /standard_name="SpCas9 sgRNA"
     misc_feature    complement(4459..4477)
                     /note="Geneious type: primer"
                     /standard_name="T7 Terminator Primer"
     regulatory      4473..4520
                     /note="Geneious type: terminator"
                     /standard_name="T7 Terminator"
     misc_feature    4702
                     /note="Geneious type: Editing History Insertion"
                     /standard_name
     CDS             complement(4741..5400)
                     /standard_name="CAT"
     rep_origin      5762..6674
     CDS             complement(6798..7880)
                     /note="lacI gene"
                     /standard_name="lacI"
     misc_feature    7961..7980
                     /note="Geneious type: primer"
                     /standard_name="ACYCDuetUP1"
     regulatory      8004..8020
                     /note="Geneious type: promoter"
                     /standard_name="T7 Promoter"
ORIGIN      
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggataaaa agtattctat tggtttagac atcggcacta attccgttgg
      121 atgggctgtc ataaccgatg aatacaaagt accttcaaag aaatttaagg tgttggggaa
      181 cacagaccgt cattcgatta aaaagaatct tatcggtgcc ctcctattcg atagtggcga
      241 aacggcagag gcgactcgcc tgaaacgaac cgctcggaga aggtatacac gtcgcaagaa
      301 ccgaatatgt tacttacaag aaatttttag caatgagatg gccaaagttg acgattcttt
      361 ctttcaccgt ttggaagagt ccttccttgt cgaagaggac aagaaacatg aacggcaccc
      421 catctttgga aacatagtag atgaggtggc atatcatgaa aagtacccaa cgatttatca
      481 cctcagaaaa aagctagttg actcaactga taaagcggac ctgaggttaa tctacttggc
      541 tcttgcccat atgataaagt tccgtgggca ctttctcatt gagggtgatc taaatccgga
      601 caactcggat gtcgacaaac tgttcatcca gttagtacaa acctataatc agttgtttga
      661 agagaaccct ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg cccgcctctc
      721 taaatcccga cggctagaaa acctgatcgc acaattaccc ggagagaaga aaaatgggtt
      781 gttcggtaac cttatagcgc tctcactagg cctgacacca aattttaagt cgaacttcga
      841 cttagctgaa gatgccaaat tgcagcttag taaggacacg tacgatgacg atctcgacaa
      901 tctactggca caaattggag atcagtatgc ggacttattt ttggctgcca aaaaccttag
      961 cgatgcaatc ctcctatctg acatactgag agttaatact gagattacca aggcgccgtt
     1021 atccgcttca atgatcaaaa ggtacgatga acatcaccaa gacttgacac ttctcaaggc
     1081 cctagtccgt cagcaactgc ctgagaaata taaggaaata ttctttgatc agtcgaaaaa
     1141 cgggtacgca ggttatattg acggcggagc gagtcaagag gaattctaca agtttatcaa
     1201 acccatatta gagaagatgg atgggacgga agagttgctt gtaaaactca atcgcgaaga
     1261 tctactgcga aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa tccacttagg
     1321 cgaattgcat gctatactta gaaggcagga ggatttttat ccgttcctca aagacaatcg
     1381 tgaaaagatt gagaaaatcc taacctttcg cataccttac tatgtgggac ccctggcccg
     1441 agggaactct cggttcgcat ggatgacaag aaagtccgaa gaaacgatta ctccctggaa
     1501 ttttgaggaa gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccaa
     1561 ctttgacaag aatttaccga acgaaaaagt attgcctaag cacagtttac tttacgagta
     1621 tttcacagtg tacaatgaac tcacgaaagt taagtatgtc actgagggca tgcgtaaacc
     1681 cgcctttcta agcggagaac agaagaaagc aatagtagat ctgttattca agaccaaccg
     1741 caaagtgaca gttaagcaat tgaaagagga ctactttaag aaaattgaat gcttcgattc
     1801 tgtcgagatc tccggggtag aagatcgatt taatgcgtca cttggtacgt atcatgacct
     1861 cctaaagata attaaagata aggacttcct ggataacgaa gagaatgaag atatcttaga
     1921 agatatagtg ttgactctta ccctctttga agatcgggaa atgattgagg aaagactaaa
     1981 aacatacgct cacctgttcg acgataaggt tatgaaacag ttaaagaggc gtcgctatac
     2041 gggctgggga cgattgtcgc ggaaacttat caacgggata agagacaagc aaagtggtaa
     2101 aactattctc gattttctaa agagcgacgg cttcgccaat aggaacttta tgcagctgat
     2161 ccatgatgac tctttaacct tcaaagagga tatacaaaag gcacaggttt ccggacaagg
     2221 ggactcattg cacgaacata ttgcgaatct tgctggttcg ccagccatca aaaagggcat
     2281 actccagaca gtcaaagtag tggatgagct agttaaggtc atgggacgtc acaaaccgga
     2341 aaacattgta atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc aaaaaaacag
     2401 tcgagagcgg atgaagagaa tagaagaggg tattaaagaa ctgggcagcc agatcttaaa
     2461 ggagcatcct gtggaaaata cccaattgca gaacgagaaa ctttacctct attacctaca
     2521 aaatggaagg gacatgtatg ttgatcagga actggacata aaccgtttat ctgattacga
     2581 cgtcgatcac attgtacccc aatccttttt gaaggacgat tcaatcgaca ataaagtgct
     2641 tacacgctcg gataagaacc gagggaaaag tgacaatgtt ccaagcgagg aagtcgtaaa
     2701 gaaaatgaag aactattggc ggcagctcct aaatgcgaaa ctgataacgc aaagaaagtt
     2761 cgataactta actaaagctg agaggggtgg cttgtctgaa cttgacaagg ccggatttat
     2821 taaacgtcag ctcgtggaaa cccgccaaat cacaaagcat gttgcacaga tactagattc
     2881 ccgaatgaat acgaaatacg acgagaacga taagctgatt cgggaagtca aagtaatcac
     2941 tttaaagtca aaattggtgt cggacttcag aaaggatttt caattctata aagttaggga
     3001 gataaataac taccaccatg cgcacgacgc ttatcttaat gccgtcgtag ggaccgcact
     3061 cattaagaaa tacccgaagc tagaaagtga gtttgtgtat ggtgattaca aagtttatga
     3121 cgtccgtaag atgatcgcga aaagcgaaca ggagataggc aaggctacag ccaaatactt
     3181 cttttattct aacattatga atttctttaa gacggaaatc actctggcaa acggagagat
     3241 acgcaaacga cctttaattg aaaccaatgg ggagacaggt gaaatcgtat gggataaggg
     3301 ccgggacttc gcgacggtga gaaaagtttt gtccatgccc caagtcaaca tagtaaagaa
     3361 aactgaggtg cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa ggaatagtga
     3421 taagctcatc gctcgtaaaa aggactggga cccgaaaaag tacggtggct tcgatagccc
     3481 tacagttgcc tattctgtcc tagtagtggc aaaagttgag aagggaaaat ccaagaaact
     3541 gaagtcagtc aaagaattat tggggataac gattatggag cgctcgtctt ttgaaaagaa
     3601 ccccatcgac ttccttgagg cgaaaggtta caaggaagta aaaaaggatc tcataattaa
     3661 actaccaaag tatagtctgt ttgagttaga aaatggccga aaacggatgt tggctagcgc
     3721 cggagagctt caaaagggga acgaactcgc actaccgtct aaatacgtga atttcctgta
     3781 tttagcgtcc cattacgaga agttgaaagg ttcacctgaa gataacgaac agaagcaact
     3841 ttttgttgag cagcacaaac attatctcga cgaaatcata gagcaaattt cggaattcag
     3901 taagagagtc atcctagctg atgccaatct ggacaaagta ttaagcgcat acaacaagca
     3961 cagggataaa cccatacgtg agcaggcgga aaatattatc catttgttta ctcttaccaa
     4021 cctcggcgct ccagccgcat tcaagtattt tgacacaacg atagatcgca aacgatacac
     4081 ttctaccaag gaggtgctag acgcgacact gattcaccaa tccatcacgg gattatatga
     4141 aactcggata gatttgtcac agcttggggg tgacggatcc cccaagaaga agaggaaagt
     4201 ctcgagcgac tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga
     4261 tgacgatgac aagtgaagcg gccgcataat gcttaagtcg aacagaaagt aatcgtattg
     4321 tacacggccg cataatcgaa attaatacga ctcactatag ggggcacggg cagcttgccg
     4381 ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     4441 tggcaccgag tcggtgctcc gctgagcaat aactagcata accccttggg gcctctaaac
     4501 gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc
     4561 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     4621 gccctgaacc gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt
     4681 atcacttatt caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa
     4741 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     4801 atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     4861 gccttgcgta taatatttgc ccatagtgaa aacgggggcg aagaagttgt ccatattggc
     4921 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     4981 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     5041 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     5101 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     5161 accgtctttc attgccatac ggaactccgg atgagcattc atcaggcggg caagaatgtg
     5221 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     5281 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     5341 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     5401 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     5461 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
     5521 aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa
     5581 gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt
     5641 actgatttag tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct
     5701 gtccctcctg ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc
     5761 agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt
     5821 gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg
     5881 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     5941 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     6001 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     6061 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     6121 cgtttcccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt
     6181 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg
     6241 cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct
     6301 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag
     6361 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa
     6421 actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag
     6481 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag
     6541 caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa
     6601 tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     6661 cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg
     6721 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     6781 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6841 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     6901 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     6961 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     7021 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     7081 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     7141 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     7201 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     7261 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     7321 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     7381 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     7441 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     7501 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     7561 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     7621 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     7681 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     7741 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     7801 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     7861 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     7921 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     7981 atgcgactcc tgcattagga aattaatacg actcactata
//
