LOCUS       Exported                6788 bp ds-DNA     circular SYN 10-4ÔÂ-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pYPQ290 (AacCas12b)m-D570A-SRDX
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6788)
  AUTHORS   Yiping Qi Lab
  TITLE     Direct Submission
  JOURNAL   Exported 2019Äê7ÔÂ14ÈÕ from SnapGene Viewer 2.8.0
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6788
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        1..138
                     /label=Spn promoter
                     /note="Spn promoter"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     dnas_title=Spn promoter
                     vntifkey=30"
     CDS             139..1149
                     /gene="Streptomycin/Spectinomycin adenyltransferase"
                     /label=SpnR
                     /note="SpnR"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     dnas_title=SpnR
                     invitrogen=2750000
                     vntifkey=4"
     rep_origin      1216..1889
                     /gene="ori"
                     /label=pUC origin
                     /note="pUC origin"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     dnas_title=pUC origin
                     invitrogen=2530000
                     vntifkey=33"
     terminator      2319..2362
                     /label=rrnB T1 terminatior
                     /note="rrnB T1 terminatior terminator"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     dnas_title=rrnB T1 terminatior
                     vntifkey=43"
     misc_recomb     2461..2560
                     /label=attL1
                     /note="attL1"
                     /note="ApEinfo_fwdcolor=#00ff40
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     dnas_title=attL1
                     invitrogen=2670000
                     vntifkey=86"
     CDS             2579..2629
                     /codon_start=1
                     /label=N-terminal NLS
                     /note="N-terminal NLS"
                     /note="N-terminal NLS
                     ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink"
                     /translation="MAPKKKRKVGIHGVPAA"
     misc_feature    2585..2605
                     /label=SV40 NLS
                     /note="SV40 NLS"
                     /note="SV40 NLS
                     ApEinfo_fwdcolor=#ff0000
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#ff0000"
     CDS             2630..6013
                     /codon_start=1
                     /note="AacCas12b (C2c1; rice codon optimized)"
                     /translation="AVKSIKVKLRLDDMPEIRAGLWKLHKEVNAGVRYYTEWLSLLRQE
                     NLYRRSPNGDGEQECDKTAEECKAELLERLRARQVENGHRGPAGSDDELLQLARQLYEL
                     LVPQAIGAKGDAQQIARKFLSPLADKDAVGGLGIAKAGNKPRWVRMREAGEPGWEEEKE
                     KAETRKSADRTADVLRALADFGLKPLMRVYTDSEMSSVEWKPLRKGQAVRTWDRDMFQQ
                     AIERMMSWESWNQRVGQEYAKLVEQKNRFEQKNFVGQEHLVHLVNQLQQDMKEASPGLE
                     SKEQTAHYVTGRALRGSDKVFEKWGKLAPDAPFDLYDAEIKNVQRRNTRRFGSHDLFAK
                     LAEPEYQALWREDASFLTRYAVYNSILRKLNHAKMFATFTLPDATAHPIWTRFDKLGGN
                     LHQYTFLFNEFGERRHAIRFHKLLKVENGVAREVDDVTVPISMSEQLDNLLPRDPNEPI
                     ALYFRDYGAEQHFTGEFGGAKIQCRRDQLAHMHRRRGARDVYLNVSVRVQSQSEARGER
                     RPPYAAVFRLVGDNHRAFVHFDKLSDYLAEHPDDGKLGSEGLLSGLRVMSVALGLRTSA
                     SISVFRVARKDELKPNSKGRVPFFFPIKGNDNLVAVHERSQLLKLPGETESKDLRAIRE
                     ERQRTLRQLRTQLAYLRLLVRCGSEDVGRRERSWAKLIEQPVDAANHMTPDWREAFENE
                     LQKLKSLHGICSDKEWMDAVYESVRRVWRHMGKQVRDWRKDVRSGERPKIRGYAKDVVG
                     GNSIEQIEYLERQYKFLKSWSFFGKVSGQVIRAEKGSRFAITLREHIDHAKEDRLKKLA
                     DRIIMEALGYVYALDERGKGKWVAKYPPCQLILLEELSEYQFNNDRPPSENNQLMQWSH
                     RGVFQELINQAQVHDLLVGTMYAAFSSRFDARTGAPGIRCRRVPARCTQEHNPEPFPWW
                     LNKFVVEHTLDACPLRADDLIPTGEGEIFVSPFSAEEGDFHQIHADLNAAQNLQQRLWS
                     DFDISQIRLRCDWGEVDGELVLIPRLTGKRTADSYSNKVFYTNTGVTYYERERGKKRRK
                     VFAQEKLSEEEAELLVEADEAREKSVVLMRDPSGIINRGNWTRQKEFWSMVNQRIEGYL
                     VKQIRSRVPLQDSACENTGDI"
     misc_feature    2974
                     /note="Feature 20"
     misc_feature    4334..4336
                     /note="D570A"
     CDS             6014..6058
                     /codon_start=1
                     /label=Nucleoplasmin NLS
                     /note="Nucleoplasmin NLS"
                     /note="Nucleoplasmin NLS
                     ApEinfo_fwdcolor=#ff0000
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#ff0000"
                     /translation="KRPAATKKAGQAKKK"
     misc_feature    6059..6103
                     /note="5X GS linker"
     exon            6104..6211
                     /note="SRDX"
     misc_feature    6236..6513
                     /label=NOS term
                     /note="NOS term"
                     /note="ApEinfo_fwdcolor=#f5010a
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=green
                     dnas_title=NOS term
                     vntifkey=21"
     misc_feature    complement(6280..6303)
                     /label=Nos-term-R2
                     /label=PCR_primer
                     /note="Nos-term-R2"
                     /note="Nos-term-R2
                     ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink"
     misc_recomb     6535..6658
                     /label=attR5
                     /note="attR5"
                     /note="ApEinfo_fwdcolor=#00ff00
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     dnas_title=attR5
                     invitrogen=2703000
                     vntifkey=86"
     promoter        6787..6788
                     /label=Spn promoter(1)
                     /note="Spn promoter(1) promoter"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_label=Spn promoter
                     ApEinfo_revcolor=pink
                     dnas_title=Spn promoter
                     vntifkey=30"
ORIGIN
        1 tggaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
       61 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
      121 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
      181 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
      241 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
      301 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacatcatg
      361 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
      421 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
      481 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
      541 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
      601 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
      661 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
      721 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
      781 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
      841 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
      901 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
      961 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     1021 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     1081 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     1141 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2101 agtgagcgca acgcaattaa tacgcgtacc gcgagccagg aagagtttgt agaaacgcaa
     2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgctt
     2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
     2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
     2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgaattcgc ccttcaccat
     2581 ggctcctaag aagaagcgga aggttggtat tcacggggtg cctgcggctg cggtgaagtc
     2641 aataaaagtt aaactccgcc tggacgatat gccagaaatt cgggctggcc tctggaagct
     2701 tcacaaagag gttaacgctg gcgtcagata ttacacggaa tggttgtcgc tgctccggca
     2761 agaaaatctc tacagaaggt cgcccaatgg tgatggggaa caagagtgcg acaaaacggc
     2821 ggaggaatgc aaagcggaac tccttgaaag acttcgcgcg agacaagtcg aaaacggcca
     2881 tagaggcccg gccggttccg atgatgaatt gcttcagctt gcgcggcagc tttacgaatt
     2941 gctcgtgccg caagccatag gtgcaaaagg agatgcacaa caaattgcaa gaaagttcct
     3001 ctccccgctc gcagacaagg atgccgtggg aggtcttgga atcgctaaag cagggaataa
     3061 gccaagatgg gtgcggatgc gggaagcagg tgagccaggc tgggaagagg agaaggagaa
     3121 agccgaaacg aggaaatcag cggatcgcac tgcagacgtg ttgagagccc tcgcagactt
     3181 tggacttaag ccactgatgc gggtttacac ggattcagag atgtcctcgg tggaatggaa
     3241 gccgctcaga aagggtcaag ccgtgagaac gtgggaccgc gacatgttcc agcaggcaat
     3301 tgagcggatg atgtcctggg agtcttggaa ccaaagggtc gggcaagaat atgcgaaact
     3361 ggtggagcaa aaaaataggt ttgaacaaaa aaatttcgtt ggtcaagagc atctggttca
     3421 tttggttaat caacttcaac aagatatgaa agaagcatca cctggcttgg aatctaaaga
     3481 acaaacagca cactacgtta cgggtagggc gttgagggga tcggataaag ttttcgagaa
     3541 gtggggtaag ttggcccccg acgccccttt cgatctgtat gacgccgaga taaagaacgt
     3601 tcagcggagg aacactcgcc gctttggttc gcacgatctg tttgcaaaac tggccgagcc
     3661 tgagtaccag gccctttggc gggaggatgc gtcgttcctt acacgctacg cggtttataa
     3721 ttcaattctc agaaagctca atcacgcgaa gatgtttgcg actttcactc ttccagatgc
     3781 gacggcacac cctatatgga ctagatttga taagttgggg ggcaacttgc accagtatac
     3841 atttctgttc aacgaattcg gcgaacgcag gcatgcaatc aggttccata aacttttgaa
     3901 agtcgagaat ggtgttgcca gggaggttga cgatgtcaca gtgcctatct cgatgtccga
     3961 acaattggat aacttgctgc ccagagatcc gaacgaaccg attgcacttt atttcaggga
     4021 ttatggtgcc gaacaacact ttacgggtga gttcggaggg gccaagattc agtgcagacg
     4081 ggaccagctt gctcacatgc accgcaggag aggggctagg gatgtgtatt tgaacgtttc
     4141 agttcgcgtg cagtcccaat ccgaggcgcg gggggagcgc agaccaccat acgcggctgt
     4201 cttccggctg gttggcgata accatagagc gttcgtgcat ttcgataagc tgagcgatta
     4261 cctcgccgaa catcctgatg acggaaagtt ggggtcagag gggcttctgt cgggcctgag
     4321 ggtgatgtcc gtggccctgg gattgcgcac cagtgcctcg atcagcgttt ttagggtggc
     4381 caggaaagat gagttgaaac ccaactcgaa ggggagggtt ccgttctttt tccctataaa
     4441 gggcaacgat aacttggtcg cagtgcatga aaggagccaa ctgctcaaac ttcccgggga
     4501 gacagagtcc aaagatcttc gcgctataag ggaagagaga caaagaactc tccggcagct
     4561 gcgcacgcag ctcgcatacc tgcggttgct tgtccgctgc ggaagtgaag acgttggcag
     4621 gcgcgagagg tcatgggcca aattgattga gcagccggtc gacgccgcaa atcacatgac
     4681 tccggattgg agggaggctt tcgagaacga actgcagaag ttgaagagtc tgcatggcat
     4741 atgctctgac aaagagtgga tggacgcggt ttacgagtcc gtccgccggg tctggcggca
     4801 catggggaaa caagttcgcg attggagaaa ggatgttaga tccggggaaa ggccgaagat
     4861 aagaggttat gccaaagacg tggttggtgg aaattctatc gaacagatcg aatatcttga
     4921 gaggcagtac aagttcctca agagttggtc tttcttcggt aaagtctctg gacaagttat
     4981 aagagcagaa aaggggagcc ggttcgctat caccttgcgg gaacacatag accacgcaaa
     5041 agaagacaga ctgaagaagc tggcggacag aattatcatg gaagcgctgg ggtacgttta
     5101 cgcgctggac gaaaggggga aaggtaaatg ggtggccaaa tacccgccat gccagttgat
     5161 attgctggaa gaattgtccg aatatcaatt taataacgat agaccgccat ccgagaacaa
     5221 ccaacttatg caatggtctc accggggagt tttccaggag ttgatcaacc aagctcaagt
     5281 gcacgatctg cttgttggta caatgtacgc agcgttttcc tcacgcttcg acgctagaac
     5341 aggagcgccg ggaattcggt gccggagggt gcctgcgagg tgtactcagg agcacaaccc
     5401 ggagccattt ccctggtggt tgaataaatt cgttgtggaa catacgttgg atgcttgccc
     5461 gcttcgggcg gacgacctca ttccgacggg tgagggcgag attttcgtgt cgccattctc
     5521 ggctgaggaa ggggacttcc atcaaatcca tgctgacctc aatgcggcgc aaaatctgca
     5581 gcagagattg tggagtgatt ttgacatctc tcagatcagg cttcggtgcg attggggaga
     5641 agtcgatggt gaactcgttc tcattccgag actcaccggt aaaaggactg ctgattcata
     5701 ttcgaacaaa gttttttaca ctaacacagg ggtcacttat tatgaaagag aacgcggtaa
     5761 gaagcgccgc aaggtgttcg cgcaagagaa actttccgag gaagaggccg agttgctcgt
     5821 tgaagctgac gaagctcgcg agaagtccgt cgttctgatg cgggatcctt ctggcataat
     5881 aaacaggggg aattggacac ggcagaagga attttggtcc atggtgaatc agcgcataga
     5941 aggttatctg gtcaaacaga tcagaagcag ggttcccctc caggattcag cgtgcgagaa
     6001 cacgggcgat attaagcgtc ctgctgccac caaaaaggcc ggacaggcta agaaaaagaa
     6061 gggagacggc tctggatcgg ggtcgggttc tggctcagtc gaccttgatc ttgacctcga
     6121 actcagactt ggatttgctc tcgatctcga ccttgaactt agactcggat ttgctcttga
     6181 cctcgatctt gagcttagac tcggattcgc ttaggacgtc ggcggccgcc gacgtccgat
     6241 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     6301 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     6361 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     6421 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     6481 ttactagatc gggaattgat cccccctcga cagcttccgg aaagggcgaa ttcgcaactt
     6541 tgtatacaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
     6601 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatgcc
     6661 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     6721 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     6781 gcctcctc
//
