LOCUS       P789_pY026_EnAsCpf1_CLYBL_T1_catRNA        9861 bp    DNA     circular UNA 23-FEB-2024
DEFINITION  EnAsCas12a (E174R/N282A/S542R/K548R) with gRNA for targeting the
            CLYBL safe harbor site.
ACCESSION   urn.local...6y-gahlkb8
VERSION     urn.local...6y-gahlkb8
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     regulatory      168..418
                     /note="Geneious type: promoter"
                     /standard_name="U6 promoter; U6 promoter (Kunkel, 1998)"
     misc_feature    420..488
                     /note="Geneious type: cat RNA"
                     /standard_name="tRNA"
     misc_feature    489..508
                     /note="Geneious type: AsCpf1 crRNA"
                     /standard_name="AsCpf1 crRNA"
     gene            509..532
                     /gene="CLYBL"
                     /gene_synonym="CLB"
                     /note="citrate lyase beta like; Derived by automated
                     computational analysis using gene prediction method:
                     BestRefSeq,Gnomon."
                     /db_xref="GeneID:171425"
                     /db_xref="HGNC:HGNC:18355"
                     /db_xref="MIM:609686"
                     /standard_name="CLYBL gene"
     misc_feature    533..552
                     /note="Geneious type: AsCpf1 crRNA"
                     /standard_name="AsCpf1 crRNA"
     regulatory      553..557
                     /note="Geneious type: terminator"
                     /standard_name="terminator"
     regulatory      626..1227
                     /note="Geneious type: promoter"
                     /standard_name="CMV promoter"
     regulatory      707..1011
                     /note="Geneious type: enhancer"
                     /note="<html><body>human cytomegalovirus immediate early
                     enhancer</body></html>"
                     /standard_name="CMV enhancer"
     regulatory      1012..1215
                     /note="Geneious type: promoter"
                     /note="<html><body>human cytomegalovirus (CMV) immediate
                     early promoter</body></html>"
                     /standard_name="CMV promoter"
     regulatory      1260..1277
                     /note="Geneious type: promoter"
                     /note="T7 RNA polymerase promoter"
                     /regulatory_class="promoter"
                     /note="Derived using Geneious Prime 2020.1.0 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="T7"
     CDS             1326..5384
                     /standard_name="huAsCpf1"
     misc_feature    1326..5384
                     /note="Geneious type: ORF"
                     /standard_name="ORF 5 (frame 3)"
     misc_feature    1845..1847
                     /note="Geneious type: mutation"
                     /standard_name="E174R"
     misc_feature    2169..2171
                     /note="Geneious type: mutation"
                     /standard_name="N282A"
     misc_feature    2949..2951
                     /note="Geneious type: mutation"
                     /standard_name="S542R"
     misc_feature    2967..2969
                     /note="Geneious type: mutation"
                     /standard_name="K548R"
     misc_feature    5247..5294
                     /note="Geneious type: HA tag"
                     /standard_name="nucleoplasmin bipartite cNLS"
     misc_feature    5301..5327
                     /standard_name="HA tag"
     misc_feature    5328..5354
                     /standard_name="HA tag"
     misc_feature    5355..5381
                     /standard_name="HA tag"
     regulatory      5458..5685
                     /note="Geneious type: Terminator"
                     /standard_name="bGH_PA term"
     misc_feature    5748..6054
                     /note="Geneious type: Origin of Replication"
                     /standard_name="f1 origin"
     regulatory      6174..6389
                     /note="Geneious type: enhancer"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="SV40 enhancer"
     misc_feature    6570..7363
                     /note="Geneious type: antibiotic resitance"
                     /standard_name="Kanamycin res"
     polyA_site      7535..7665
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
     misc_feature    complement(7725..7752)
                     /note="Geneious type: Regulatory Sequence"
                     /standard_name="lacO reg"
     regulatory      complement(7756..7785)
                     /note="Geneious type: Promoter"
                     /standard_name="lac prom"
     misc_feature    complement(7800..7821)
                     /note="Geneious type: CAP"
                     /standard_name="CAP binding site"
     misc_RNA        complement(8109..8658)
                     /note="RNAII transcript (complementary strand)"
                     /standard_name="RNA II; RNA II; RNA"
     rep_origin      complement(8109..8694)
     misc_RNA        8551..8655
                     /note="Geneious type: misc_rna"
                     /note="RNAI transcript"
                     /standard_name="RNAI transcript"
     misc_feature    complement(8865..9725)
                     /note="Geneious type: Marker"
                     /standard_name="amp marker"
     regulatory      complement(9726..9817)
                     /note="Geneious type: promoter"
                     /gene="<html><body><i>bla</i></body></html>"
                     /standard_name="AmpR promoter"
     regulatory      complement(9767..9795)
                     /note="Geneious type: Promoter"
                     /standard_name="amp prom"
ORIGIN      
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga cttaattaag agggcctatt
      181 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
      241 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
      301 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
      361 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
      421 ccagtggtct agtggtagaa tagtaccctg ccacggtaca gacccgggtt cgattcccgg
      481 ctggaaatta atttctactc ttgtagatac ttccttactg ttaacttcca tataatttct
      541 actcttgtag attttttcaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
      601 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
      661 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      721 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      781 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      841 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      901 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      961 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1021 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     1081 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     1141 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     1201 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     1261 aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg
     1321 ccaccatgac acagttcgag ggctttacca acctgtatca ggtgagcaag acactgcggt
     1381 ttgagctgat cccacagggc aagaccctga agcacatcca ggagcagggc ttcatcgagg
     1441 aggacaaggc ccgcaatgat cactacaagg agctgaagcc catcatcgat cggatctaca
     1501 agacctatgc cgaccagtgc ctgcagctgg tgcagctgga ttgggagaac ctgagcgccg
     1561 ccatcgactc ctatagaaag gagaaaaccg aggagacaag gaacgccctg atcgaggagc
     1621 aggccacata tcgcaatgcc atccacgact acttcatcgg ccggacagac aacctgaccg
     1681 atgccatcaa taagagacac gccgagatct acaagggcct gttcaaggcc gagctgttta
     1741 atggcaaggt gctgaagcag ctgggcaccg tgaccacaac cgagcacgag aacgccctgc
     1801 tgcggagctt cgacaagttt acaacctact tctccggctt ttataggaac aggaagaacg
     1861 tgttcagcgc cgaggatatc agcacagcca tcccacaccg catcgtgcag gacaacttcc
     1921 ccaagtttaa ggagaattgt cacatcttca cacgcctgat caccgccgtg cccagcctgc
     1981 gggagcactt tgagaacgtg aagaaggcca tcggcatctt cgtgagcacc tccatcgagg
     2041 aggtgttttc cttccctttt tataaccagc tgctgacaca gacccagatc gacctgtata
     2101 accagctgct gggaggaatc tctcgggagg caggcaccga gaagatcaag ggcctgaacg
     2161 aggtgctggc cctggccatc cagaagaatg atgagacagc ccacatcatc gcctccctgc
     2221 cacacagatt catccccctg tttaagcaga tcctgtccga taggaacacc ctgtctttca
     2281 tcctggagga gtttaagagc gacgaggaag tgatccagtc cttctgcaag tacaagacac
     2341 tgctgagaaa cgagaacgtg ctggagacag ccgaggccct gtttaacgag ctgaacagca
     2401 tcgacctgac acacatcttc atcagccaca agaagctgga gacaatcagc agcgccctgt
     2461 gcgaccactg ggatacactg aggaatgccc tgtatgagcg gagaatctcc gagctgacag
     2521 gcaagatcac caagtctgcc aaggagaagg tgcagcgcag cctgaagcac gaggatatca
     2581 acctgcagga gatcatctct gccgcaggca aggagctgag cgaggccttc aagcagaaaa
     2641 ccagcgagat cctgtcccac gcacacgccg ccctggatca gccactgcct acaaccctga
     2701 agaagcagga ggagaaggag atcctgaagt ctcagctgga cagcctgctg ggcctgtacc
     2761 acctgctgga ctggtttgcc gtggatgagt ccaacgaggt ggaccccgag ttctctgccc
     2821 ggctgaccgg catcaagctg gagatggagc cttctctgag cttctacaac aaggccagaa
     2881 attatgccac caagaagccc tactccgtgg agaagttcaa gctgaacttt cagatgccta
     2941 cactggccag gggctgggac gtgaataggg agaagaacaa tggcgccatc ctgtttgtga
     3001 agaacggcct gtactatctg ggcatcatgc caaagcagaa gggcaggtat aaggccctga
     3061 gcttcgagcc cacagagaaa accagcgagg gctttgataa gatgtactat gactacttcc
     3121 ctgatgccgc caagatgatc ccaaagtgca gcacccagct gaaggccgtg acagcccact
     3181 ttcagaccca cacaaccccc atcctgctgt ccaacaattt catcgagcct ctggagatca
     3241 caaaggagat ctacgacctg aacaatcctg agaaggagcc aaagaagttt cagacagcct
     3301 acgccaagaa aaccggcgac cagaagggct acagagaggc cctgtgcaag tggatcgact
     3361 tcacaaggga ttttctgtcc aagtatacca agacaacctc tatcgatctg tctagcctgc
     3421 ggccatcctc tcagtataag gacctgggcg agtactatgc cgagctgaat cccctgctgt
     3481 accacatcag cttccagaga atcgccgaga aggagatcat ggatgccgtg gagacaggca
     3541 agctgtacct gttccagatc tataacaagg actttgccaa gggccaccac ggcaagccta
     3601 atctgcacac actgtattgg accggcctgt tttctccaga gaacctggcc aagacaagca
     3661 tcaagctgaa tggccaggcc gagctgttct accgccctaa gtccaggatg aagaggatgg
     3721 cacaccggct gggagagaag atgctgaaca agaagctgaa ggatcagaaa accccaatcc
     3781 ccgacaccct gtaccaggag ctgtacgact atgtgaatca cagactgtcc cacgacctgt
     3841 ctgatgaggc cagggccctg ctgcccaacg tgatcaccaa ggaggtgtct cacgagatca
     3901 tcaaggatag gcgctttacc agcgacaagt tctttttcca cgtgcctatc acactgaact
     3961 atcaggccgc caattcccca tctaagttca accagagggt gaatgcctac ctgaaggagc
     4021 accccgagac acctatcatc ggcatcgatc ggggcgagag aaacctgatc tatatcacag
     4081 tgatcgactc caccggcaag atcctggagc agcggagcct gaacaccatc cagcagtttg
     4141 attaccagaa gaagctggac aacagggaga aggagagggt ggcagcaagg caggcctggt
     4201 ctgtggtggg cacaatcaag gatctgaagc agggctatct gagccaggtc atccacgaga
     4261 tcgtggacct gatgatccac taccaggccg tggtggtgct ggagaacctg aatttcggct
     4321 ttaagagcaa gaggaccggc atcgccgaga aggccgtgta ccagcagttc gagaagatgc
     4381 tgatcgataa gctgaattgc ctggtgctga aggactatcc agcagagaaa gtgggaggcg
     4441 tgctgaaccc ataccagctg acagaccagt tcacctcctt tgccaagatg ggcacccagt
     4501 ctggcttcct gttttacgtg cctgccccat atacatctaa gatcgatccc ctgaccggct
     4561 tcgtggaccc cttcgtgtgg aaaaccatca agaatcacga gagccgcaag cacttcctgg
     4621 agggcttcga ctttctgcac tacgacgtga aaaccggcga cttcatcctg cactttaaga
     4681 tgaacagaaa tctgtccttc cagaggggcc tgcccggctt tatgcctgca tgggatatcg
     4741 tgttcgagaa gaacgagaca cagtttgacg ccaagggcac ccctttcatc gccggcaaga
     4801 gaatcgtgcc agtgatcgag aatcacagat tcaccggcag ataccgggac ctgtatcctg
     4861 ccaacgagct gatcgccctg ctggaggaga agggcatcgt gttcagggat ggctccaaca
     4921 tcctgccaaa gctgctggag aatgacgatt ctcacgccat cgacaccatg gtggccctga
     4981 tccgcagcgt gctgcagatg cggaactcca atgccgccac aggcgaggac tatatcaaca
     5041 gccccgtgcg cgatctgaat ggcgtgtgct tcgactcccg gtttcagaac ccagagtggc
     5101 ccatggacgc cgatgccaat ggcgcctacc acatcgccct gaagggccag ctgctgctga
     5161 atcacctgaa ggagagcaag gatctgaagc tgcagaacgg catctccaat caggactggc
     5221 tggcctacat ccaggagctg cgcaacaaaa ggccggcggc cacgaaaaag gccggccagg
     5281 caaaaaagaa aaagggatcc tacccatacg atgttccaga ttacgcttat ccctacgacg
     5341 tgcctgatta tgcataccca tatgatgtcc ccgactatgc ctaagaattc tgcagatatc
     5401 cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     5461 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5521 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5581 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5641 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     5701 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     5761 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5821 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5881 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5941 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6001 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6061 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6121 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     6181 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6241 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6301 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6361 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6421 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     6481 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     6541 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     6601 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     6661 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     6721 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     6781 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     6841 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     6901 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6961 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     7021 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     7081 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     7141 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     7201 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     7261 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     7321 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     7381 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     7441 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     7501 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     7561 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     7621 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     7681 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     7741 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     7801 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7861 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7921 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7981 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     8041 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     8101 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     8161 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     8221 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     8281 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     8341 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     8401 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     8461 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     8521 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     8581 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     8641 agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     8701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     8761 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     8821 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     8881 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     8941 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     9001 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     9061 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     9121 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     9181 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     9241 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     9301 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     9361 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     9421 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     9481 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     9541 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     9601 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     9661 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     9721 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     9781 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     9841 cgaaaagtgc cacctgacgt c
//
