LOCUS       TwinStrepSUMO-BhCas12bv4-NLS        9619 bp ds-DNA     circular     14-JUL-2020
DEFINITION  .
ACCESSION   urn.local...y-c5uzmhh
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     source          1..9619
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     310..329
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /label="T7"
     misc_binding    329..351
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /label="LacO"
     misc_feature    397..549
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /label="6x His tag/ Twin Strep"
     misc_feature    550..841
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /label="Sumo"
     misc_feature    847..897
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /label="NLS"
     misc_feature    898..4218
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /label="BhC2C1"
     misc_feature    4219..4266
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /label="NLS nuc"
     misc_feature    4267..4353
                     /created_by="Jonathan Strecker"
                     /label="3xHA"
     rep_origin      complement(7774..8456)
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /label="ColE1 origin"
     CDS             complement(8554..9213)
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="AmpR"
ORIGIN      
        1 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
       61 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
      121 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
      181 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
      241 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
      301 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
      361 aaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat
      421 catcacagca gcggcctggt gccgcgcggc agccatatgg ctagctggag ccatccgcag
      481 tttgaaaaag gtggtggtag cggtggtggt tcaggtggta gtgcatggtc acaccctcag
      541 tttgagaaaa tgtcggactc agaagtcaat caagaagcta agccagaggt caagccagaa
      601 gtcaagcctg agactcacat caatttaaag gtgtccgatg gatcttcaga gatcttcttc
      661 aagatcaaaa agaccactcc tttaagaagg ctgatggaag cgttcgctaa aagacagggt
      721 aaggaaatgg actccttaag attcttgtac gacggtatta gaatccaagc tgatcagacc
      781 cctgaagatt tggacatgga ggataacgat attattgagg ctcacagaga acagattggt
      841 ggatccatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgcc
      901 accagatcct tcatcctgaa gatcgagccc aacgaggaag tgaagaaagg cctctggaaa
      961 acccacgagg tgctgaacca cggaatcgcc tactacatga atatcctgaa gctgatccgg
     1021 caagaggcca tctacgagca ccacgagcag gaccccaaga atcccaagaa ggtgtccaag
     1081 gccgagatcc aggccgagct gtgggatttc gtgctgaaga tgcagaagtg caacagcttc
     1141 acacacgagg tggacaagga cgaggtgttc aacatcctga gagagctgta cgaggaactg
     1201 gtgcccagca gcgtggaaaa gaagggcgaa gccaaccagc tgagcaacaa gtttctgtac
     1261 cctctggtgg accccaacag ccagtctgga aagggaacag ccagcagcgg cagaaagccc
     1321 agatggtaca acctgaagat tgccggcgat ccctcctggg aagaagagaa gaagaagtgg
     1381 gaagaagata agaaaaagga cccgctggcc aagatcctgg gcaagctggc tgagtacgga
     1441 ctgatccctc tgttcatccc ctacaccgac agcaacgagc ccatcgtgaa agaaatcaag
     1501 tggatggaaa agtcccggaa ccagagcgtg cggcggctgg ataaggacat gttcattcag
     1561 gccctggaac ggttcctgag ctgggagagc tggaacctga aagtgaaaga ggaatacgag
     1621 aaggtcgaga aagagtacaa gaccctggaa gagaggatca aagaggacat ccaggctctg
     1681 aaggctctgg aacagtatga gaaagagcgg caagaacagc tgctgcggga caccctgaac
     1741 accaacgagt accggctgag caagagaggc cttagaggct ggcgggaaat catccagaaa
     1801 tggctgaaaa tggacgagaa cgagccctcc gagaagtacc tggaagtgtt caaggactac
     1861 cagcggaagc accctagaga ggccggcgat tacagcgtgt acgagttcct gtccaagaaa
     1921 gagaaccact tcatctggcg gaatcaccct gagtacccct acctgtacgc caccttctgc
     1981 gagatcgaca agaaaaagaa ggacgccaag cagcaggcca ccttcacact ggccgatcct
     2041 atcaatcacc ctctgtgggt ccgattcgag gaaagaagcg gcagcaacct gaacaagtac
     2101 agaatcctga ccgagcagct gcacaccgag aagctgaaga aaaagctgac agtgcagctg
     2161 gaccggctga tctaccctac agaatctggc ggctgggaag agaagggcaa agtggacatt
     2221 gtgctgctgc ccagccggca gttctacaac cagatcttcc tggacatcga ggaaaagggc
     2281 aagcacgcct tcacctacaa ggatgagagc atcaagttcc ctctgaaggg cacactcggc
     2341 ggagccagag tgcagttcga cagagatcac ctgagaagat accctcacaa ggtggaaagc
     2401 ggcaacgtgg gcagaatcta cttcaacatg accgtgaaca tcgagcctac agagtcccca
     2461 gtgtccaagt ctctgaagat ccaccgggac gacttcccca aggtggtcaa cttcaagccc
     2521 aaagaactga ccgagtggat caaggacagc aagggcaaga aactgaagtc cggcatcgag
     2581 tccctggaaa tcggcctgag agtgatgagc atcgacctgg gacagagaca ggccgctgcc
     2641 gcctctattt tcgaggtggt ggatcagaag cccgacatcg aaggcaagct gtttttccca
     2701 atcaagggca ccgagctgta tgccgtgcac agagccagct tcaacatcaa gctgcccggc
     2761 gagacactgg tcaagagcag agaagtgctg cggaaggcca gagaggacaa tctgaaactg
     2821 atgaaccaga agctcaactt cctgcggaac gtgctgcact tccagcagtt cgaggacatc
     2881 accgagagag agaagcgggt caccaagtgg atcagcagac aagagaacag cgacgtgccc
     2941 ctggtgtacc aggatgagct gatccagatc cgcgagctga tgtacaagcc ttacaaggac
     3001 tgggtcgcct tcctgaagca gctccacaag agactggaag tcgagatcgg caaagaagtg
     3061 aagcactggc ggaagtccct gagcgacgga agaaagggcc tgtacggcat ctccctgaag
     3121 aacatcgacg agatcgatcg gacccggaag ttcctgctga gatggtccct gaggcctacc
     3181 gaacctggcg aagtgcgtag actggaaccc ggccagagat tcgccatcga ccagctgaat
     3241 cacctgaacg ccctgaaaga agatcggctg aagaagatgg ccaacaccat catcatgcac
     3301 gccctgggct actgctacga cgtgcggaag aagaaatggc aggctaagaa ccccgcctgc
     3361 cagatcatcc tgttcgagga tctgagcaac tacaacccct acggagaaag gtcccgcttc
     3421 gagaacagcc ggctcatgaa gtggtccaga cgcgagatcc ccagacaggt tgcactgcag
     3481 ggcgagatct atggcctgca agtgggagaa gtgggcgctc agttcagcag cagattccac
     3541 gccaagacag gcagccctgg catcagatgt cgggtcgtga ccaaagagaa gctgcaggac
     3601 aatcggttct tcaagaatct gcagagagag ggcagactga ccctggacaa aatcgccgtg
     3661 ctgaaagagg gcgatctgta cccagacaaa ggcggcgaga agttcatcag cctgagcaag
     3721 gatcggaagt gcgtgaccac acacgccgac atcaacgccg ctcagaacct gcagaagcgg
     3781 ttctggacaa gaacccacgg cttctacaag gtgtactgca aggcctacca ggtggacggc
     3841 cagaccgtgt acatccctga gagcaaggac cagaagcaga agatcatcga agagttcggc
     3901 gagggctact tcattctgaa ggacggggtg tacgaatggg tcaacgccgg caagctgaaa
     3961 atcaagaagg gcagctccaa gcagagcagc agcgagctgg tggatagcga catcctgaaa
     4021 gacagcttcg acctggcctc cgagctgaaa ggcgaaaagc tgatgctgta cagggacccc
     4081 agcggcaatg tgttccccag cgacaaatgg atggccgctg gcgtgttctt cggaaagctg
     4141 gaacgcatcc tgatcagcaa gctgaccaac cagtactcca tcagcaccat cgaggacgac
     4201 agcagcaagc agtctatgaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     4261 aaaaagggat cctacccata cgatgttcca gattacgctt atccctacga cgtgcctgat
     4321 tatgcatacc catatgatgt ccccgactat gcctaagaat tctgcagata tccagcacag
     4381 tggcggccgc actcgaggcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     4441 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     4501 gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     4561 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     4621 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     4681 gataagctgt caaacatgag aaacaccatc gaatggcgca aaacctttcg cggtatggca
     4741 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     4801 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     4861 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     4921 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     4981 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     5041 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     5101 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     5161 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     5221 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     5281 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     5341 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     5401 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     5461 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     5521 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     5581 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
     5641 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     5701 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     5761 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     5821 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     5881 aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc
     5941 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     6001 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     6061 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
     6121 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     6181 cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg
     6241 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     6301 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     6361 gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta
     6421 tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct
     6481 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga
     6541 agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg
     6601 cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc
     6661 tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg
     6721 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
     6781 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
     6841 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     6901 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
     6961 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
     7021 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
     7081 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
     7141 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
     7201 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
     7261 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
     7321 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     7381 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     7441 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     7501 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccattgcg
     7561 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     7621 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7681 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7741 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7801 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7861 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7921 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7981 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     8041 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     8101 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8161 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8221 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8281 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8341 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8401 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8461 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8521 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8581 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8641 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8701 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8761 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8821 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt
     8881 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8941 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     9001 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9061 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9121 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     9181 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9241 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9301 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9361 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9421 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9481 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9541 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     9601 gccctttcgt cttcaagaa
//
