LOCUS       Exported                6579 bp ds-DNA     circular SYN 28-2ÔÂ-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pYPQ291 (BthCas12b)-D573A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6579)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2019Äê7ÔÂ14ÈÕ from SnapGene Viewer 2.8.0
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6579
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1..2629
                     /label=source
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink
                     dnas_title=pYPQ230 (LbCpf1)"
     source          2578..2629
                     /label=source(2)
                     /note="source(2)"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_label=source
                     ApEinfo_revcolor=pink
                     dnas_title=LbCpf1"
     source          2578..2627
                     /label=source(3)
                     /note="source(3)"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_label=source
                     ApEinfo_revcolor=pink
                     dnas_title=AsCpf1"
     source          5951..6579
                     /label=source(4)
                     /note="source(4)"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_label=source
                     ApEinfo_revcolor=pink
                     dnas_title=pYPQ230 (LbCpf1)"
     source          5951..6021
                     /label=source(1)
                     /note="source(1)"
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_label=source
                     ApEinfo_revcolor=pink
                     dnas_title=AsCpf1"
     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..5950
                     /codon_start=1
                     /note="BthCas12b (C2c1; rice codon optimized)"
                     /translation="ATRSFILKIEPNEEVKKGLWKTHEVLNHGIAYYMNILKLIRQEAI
                     YEHHEQDPKNPKKVSKAEIQAELWDFVLKMQKCNSFTHEVDKDVVFNILRELYEELVPS
                     SVEKKGEANQLSNKFLYPLVDPNSQSGKGTASSGRKPRWYNLKIAGDPSWEEEKKKWEE
                     DKKKDPLAKILGKLAEYGLIPLFIPFTDSNEPIVKEIKWMEKSRNQSVRRLDKDMFIQA
                     LERFLSWESWNLKVKEEYEKVEKEHKTLEERIKEDIQAFKSLEQYEKERQEQLLRDTLN
                     TNEYRLSKRGLRGWREIIQKWLKMDENEPSEKYLEVFKDYQRKHPREAGDYSVYEFLSK
                     KENHFIWRNHPEYPYLYATFCEIDKKKKDAKQQATFTLADPINHPLWVRFEERSGSNLN
                     KYRILTEQLHTEKLKKKLTVQLDRLIYPTESGGWEEKGKVDIVLLPSRQFYNQIFLDIE
                     EKGKHAFTYKDESIKFPLKGTLGGARVQFDRDHLRRYPHKVESGNVGRIYFNMTVNIEP
                     TESPVSKSLKIHRDDFPKFVNFKPKELTEWIKDSKGKKLKSGIESLEIGLRVMSIALGQ
                     RQAAAASIFEVVDQKPDIEGKLFFPIKGTELYAVHRASFNIKLPGETLVKSREVLRKAR
                     EDNLKLMNQKLNFLRNVLHFQQFEDITEREKRVTKWISRQENSDVPLVYQDELIQIREL
                     MYKPYKDWVAFLKQLHKRLEVEIGKEVKHWRKSLSDGRKGLYGISLKNIDEIDRTRKFL
                     LRWSLRPTEPGEVRRLEPGQRFAIDQLNHLNALKEDRLKKMANTIIMHALGYCYDVRKK
                     KWQAKNPACQIILFEDLSNYNPYEERSRFENSKLMKWSRREIPRQVALQGEIYGLQVGE
                     VGAQFSSRFHAKTGSPGIRCSVVTKEKLQDNRFFKNLQREGRLTLDKIAVLKEGDLYPD
                     KGGEKFISLSKDRKLVTTHADINAAQNLQKRFWTRTHGFYKVYCKAYQVDGQTVYIPES
                     KDQKQKIIEEFGEGYFILKDGVYEWGNAGKLKIKKGSSKQSSSELVDSDILKDSFDLAS
                     ELKGEKLMLYRDPSGNVFPSDKWMAAGVFFGKLERILISKLTNQYSISTIEDDSSKQSM
                     "
     misc_feature    4346..4348
                     /note="D573A"
     CDS             5951..5998
                     /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="KRPAATKKAGQAKKKK"
     misc_feature    5999..6001
                     /label=stop
                     /note="stop"
                     /note="stop
                     ApEinfo_fwdcolor=#ff0000
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#ff0000"
     misc_feature    6027..6304
                     /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(6071..6094)
                     /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     6326..6449
                     /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        6578..6579
                     /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 ccacaaggtc
     2641 tttcatactt aagatagagc caaacgaaga ggtcaaaaag ggattgtgga aaacccatga
     2701 agtcctgaac catggcattg cctactacat gaacatcctg aaacttatac ggcaggaggc
     2761 tatttatgag caccacgagc aggatccaaa aaaccccaaa aaggtttcga aggctgaaat
     2821 ccaggccgaa ctgtgggact tcgttctcaa aatgcagaaa tgtaattcgt tcactcatga
     2881 agttgacaaa gacgtcgtgt ttaacatttt gagggagctt tacgaggagt tggttccgag
     2941 ctccgtcgaa aagaagggtg aagcaaatca gctgtcgaat aagttcttgt accctttggt
     3001 ggacccgaac agccaatctg gaaaagggac agcatcatca gggcggaagc ctcggtggta
     3061 taacttgaag attgctggag acccttcgtg ggaagaggaa aagaaaaagt gggaggaaga
     3121 taagaagaag gacccacttg ccaaaattct cggcaaactt gccgaatatg gattgatacc
     3181 gctgttcatc ccctttacgg attctaacga acccatcgtt aaagaaatca agtggatgga
     3241 aaaatctcgc aatcagtccg tccggaggct ggacaaagat atgtttatac aagctttgga
     3301 acgctttctc tcgtgggagt cgtggaatct taaggtcaaa gaagagtatg aaaaggtcga
     3361 gaaggaacac aagacactgg aggagaggat taaggaagac attcaagcat tcaagtcact
     3421 ggagcaatac gaaaaggaac ggcaggagca attgcttcgc gacacgctca ataccaatga
     3481 atataggctt tccaagaggg gcctgagagg atggcgggaa ataatccaga aatggctcaa
     3541 gatggacgag aatgaacctt cagaaaaata tctcgaggtt tttaaagatt accaaaggaa
     3601 acatccacgc gaggcagggg attacagcgt gtacgagttt ctctccaaga aggaaaacca
     3661 ttttatctgg cgcaatcatc ccgaataccc gtacctctat gcgacgttct gcgaaataga
     3721 caaaaagaaa aaagatgcta agcaacaagc gactttcaca cttgcagatc ccataaatca
     3781 cccattgtgg gtgcggtttg aagaaaggtc gggctctaac ctcaataagt acagaatttt
     3841 gacggagcag ttgcacacag aaaagctgaa gaagaagttg acggttcagc tggatcgcct
     3901 tatctaccca accgagtctg gtggctggga agagaagggg aaagtcgaca tagtgttgct
     3961 gccatctagg cagttctata accagatttt tctcgatata gaagaaaagg gtaaacatgc
     4021 atttacgtat aaagacgagt ccataaagtt tccactgaaa ggaacacttg gcggcgcaag
     4081 ggtgcagttt gatcgggacc accttcgcag gtacccccac aaggttgaaa gtggaaacgt
     4141 tggacggatc tattttaata tgaccgtcaa catagaaccc acagaatccc ctgtttccaa
     4201 atccctgaaa atacaccggg acgattttcc taaatttgtg aactttaaac cgaaggagtt
     4261 gaccgagtgg ataaaggaca gtaaagggaa aaagctgaag tccggtatcg aaagcctgga
     4321 gattgggctc agagttatgt cgatagcgct gggtcaaagg caggcagcag ccgcctctat
     4381 atttgaggtc gtggaccaga agcccgacat tgaaggtaaa ctgttctttc cgattaaggg
     4441 gacggaactc tacgcagtcc atcgcgcctc cttcaatata aagctgccgg gcgaaacact
     4501 ggttaaatca cgcgaggttt tgcgcaaagc gcgggaagac aacctgaaac tcatgaatca
     4561 aaagctcaat ttcctgcgca atgtgttgca cttccagcag tttgaggata ttaccgaaag
     4621 agagaaaagg gttacaaaat ggatatcccg gcaagaaaac tctgatgttc cgctggttta
     4681 ccaggatgag cttatacaga ttagggaact tatgtataaa ccttacaaag attgggttgc
     4741 attcctcaag cagctgcata agagacttga agtcgagatc ggcaaagaag tcaaacactg
     4801 gcgcaagagc ctgagcgatg gtcggaaagg gttgtacgga atcagtttga aaaatatcga
     4861 cgaaatagat agaaccagga aatttttgtt gcgctggtca ctgagaccaa cggaaccggg
     4921 agaagtcaga aggttggagc caggccagag atttgcaatt gaccagctga accatctgaa
     4981 tgcactgaaa gaggacagat tgaagaagat ggcgaatacg attattatgc atgctttggg
     5041 ttattgttac gacgttagga agaagaaatg gcaggccaag aaccctgcgt gccaaatcat
     5101 cctgttcgaa gatctgagta actacaatcc gtatgaagaa aggagtcgct tcgagaacag
     5161 taaactgatg aaatggtccc ggcgcgagat accacgccaa gttgcgcttc aaggggaaat
     5221 atacgggctt caagttgggg aagttggagc gcagttttct agccggttcc acgccaagac
     5281 agggtccccg ggtataaggt gcagtgtggt gacgaaagaa aagttgcagg ataatagatt
     5341 ctttaaaaat cttcaacggg aagggcgcct gacgcttgac aagattgcag tgttgaaaga
     5401 gggggatttg taccccgata aaggcgggga gaagttcatt tctttgtcga aggaccgcaa
     5461 gttggttacg acgcatgcag acattaacgc agcacaaaat ctgcaaaaaa gattctggac
     5521 tcggacgcat ggtttttaca aggtttactg taaagcatat caagtcgatg gtcagacggt
     5581 ttacattccc gaatctaaag atcagaaaca gaaaatcatt gaggagttcg gtgaaggtta
     5641 ctttatactc aaggacggtg tttacgaatg gggtaatgct ggtaaactga aaattaagaa
     5701 ggggtcctcc aagcaatcat cttctgagct cgtcgacagc gacatcctta aggatagctt
     5761 cgatcttgcc tctgagctca agggagaaaa gttgatgctg tatcgcgatc ctagtggaaa
     5821 tgtctttccc tcagataaat ggatggcagc aggtgtgttc ttcgggaaat tggaacgcat
     5881 actgatatca aaactgacca atcaatactc tatatctact attgaagacg attcaagtaa
     5941 gcaatcgatg aagcgtcctg ctgccaccaa aaaggccgga caggctaaga aaaagaagtg
     6001 agacgactag tggcggccgc cgacgtccga tcgttcaaac atttggcaat aaagtttctt
     6061 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
     6121 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     6181 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
     6241 ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattga tcccccctcg
     6301 acagcttccg gaaagggcga attcgcaact ttgtatacaa aagttgaacg agaaacgtaa
     6361 aatgatataa atatcaatat attaaattag attttgcata aaaaacagac tacataatac
     6421 tgtaaaacac aacatatcca gtcactatgc catccagctg atatccccta tagtgagtcg
     6481 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     6541 tacattgcac aagataaaaa tatatcatca tgcctcctc
//
