LOCUS       pCMV/TO-CasRx-NLS-T2A-m 6751 bp ds-DNA     circular     02-FEB-2023
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS             complement(665..733)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        complement(734..836)
                     /label="cat promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     terminator      837..868
                     /label="tonB terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    883..1032
                     /label="stuffer 150"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          1024..1049
                     /label="pCMV/TO-CasRx-NLS-T2A-mCherry_Fw"
                     /note="sequence: ATAGATCGTCTCATGAtctggactcgttacataacttacgg"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     promoter        1033..1336
                     /label="CMV enhancer"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     promoter        1337..1540
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     TATA_signal     1526..1532
                     /label="TATA box"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     protein_bind    1542..1560
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     protein_bind    1563..1581
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     regulatory      1675..1684
                     /label="Kozak sequence"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             1696..1716
                     /label="SV40 NLS"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             1726..4623
                     /label="RfxCas13d"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             4633..4653
                     /label="SV40 NLS"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             4663..4689
                     /label="HA"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             4705..4758
                     /label="T2A"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    4765..5469
                     /label="mCherry"
                     /ApEinfo_revcolor="#dc143c"
                     /ApEinfo_fwdcolor="#dc143c"
     misc_feature    5524..5643
                     /label="SV40 poly(A)"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(5645..5673)
                     /label="pCMV/TO-CasRx-NLS-T2A-mCherry_Rv"
                     /note="sequence: ATAGATCGTCTCAacatcctcatcatagacgacagacatg"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     terminator      5923..5952
                     /label="T3Te terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     rep_origin      5974..6561
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     terminator      6573..6600
                     /label="T7Te terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             complement(6624..664)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
ORIGIN
        1 caatgatacc gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag
       61 ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta
      121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
      181 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
      301 ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta
      361 tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg
      421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
      481 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
      541 gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga
      601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
      661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
      721 gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
      781 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc
      841 aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg
      901 gtcagtattg agcgatatct agagaattcg tataggtctt gtattcagtc tcgcacttta
      961 gacatggtct tctgctgccc gtggtcctct attcgtcgga tgttctgtcg attcttcctg
     1021 tctatctgga ctcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1081 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1141 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1201 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1261 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1321 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     1381 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     1441 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     1501 cggtaggcgt gtacggtggg aggtctatat aagcagagct CTCCCTATCA GTGATAGAGA
     1561 TCTCCCTATC AGTGATAGAG ATCGTCGACG AGggacttag cctttagtga accgtcagaa
     1621 ttaattcaga tcgatctacc agaaccgtca gatccgctag agattacgcc aaccgccacc
     1681 atgggcagca tgagccccaa gaagaagaga aaggtggagg ccagcatcga aaaaaaaaag
     1741 tccttcgcca agggcatggg cgtgaagtcc acactcgtgt ccggctccaa agtgtacatg
     1801 acaaccttcg ccgaaggcag cgacgccagg ctggaaaaga tcgtggaggg cgacagcatc
     1861 aggagcgtga atgagggcga ggccttcagc gctgaaatgg ccgataaaaa cgccggctat
     1921 aagatcggca acgccaaatt cagccatcct aagggctacg ccgtggtggc taacaaccct
     1981 ctgtatacag gacccgtcca gcaggatatg ctcggcctga aggaaactct ggaaaagagg
     2041 tacttcggcg agagcgctga tggcaatgac aatatttgta tccaggtgat ccataacatc
     2101 ctggacattg aaaaaatcct cgccgaatac attaccaacg ccgcctacgc cgtcaacaat
     2161 atctccggcc tggataagga cattattgga ttcggcaagt tctccacagt gtatacctac
     2221 gacgaattca aagaccccga gcaccatagg gccgctttca acaataacga taagctcatc
     2281 aacgccatca aggcccagta tgacgagttc gacaacttcc tcgataaccc cagactcggc
     2341 tatttcggcc aggccttttt cagcaaggag ggcagaaatt acatcatcaa ttacggcaac
     2401 gaatgctatg acattctggc cctcctgagc ggactgaggc actgggtggt ccataacaac
     2461 gaagaagagt ccaggatctc caggacctgg ctctacaacc tcgataagaa cctcgacaac
     2521 gaatacatct ccaccctcaa ctacctctac gacaggatca ccaatgagct gaccaactcc
     2581 ttctccaaga actccgccgc caacgtgaac tatattgccg aaactctggg aatcaaccct
     2641 gccgaattcg ccgaacaata tttcagattc agcattatga aagagcagaa aaacctcgga
     2701 ttcaatatca ccaagctcag ggaagtgatg ctggacagga aggatatgtc cgagatcagg
     2761 aaaaatcata aggtgttcga ctccatcagg accaaggtct acaccatgat ggactttgtg
     2821 atttataggt attacatcga agaggatgcc aaggtggctg ccgccaataa gtccctcccc
     2881 gataatgaga agtccctgag cgagaaggat atctttgtga ttaacctgag gggctccttc
     2941 aacgacgacc agaaggatgc cctctactac gatgaagcta atagaatttg gagaaagctc
     3001 gaaaatatca tgcacaacat caaggaattt aggggaaaca agacaagaga gtataagaag
     3061 aaggacgccc ctagactgcc cagaatcctg cccgctggcc gtgatgtttc cgccttcagc
     3121 aaactcatgt atgccctgac catgttcctg gatggcaagg agatcaacga cctcctgacc
     3181 accctgatta ataaattcga taacatccag agcttcctga aggtgatgcc tctcatcgga
     3241 gtcaacgcta agttcgtgga ggaatacgcc tttttcaaag actccgccaa gatcgccgat
     3301 gagctgaggc tgatcaagtc cttcgctaga atgggagaac ctattgccga tgccaggagg
     3361 gccatgtata tcgacgccat ccgtatttta ggaaccaacc tgtcctatga tgagctcaag
     3421 gccctcgccg acaccttttc cctggacgag aacggaaaca agctcaagaa aggcaagcac
     3481 ggcatgagaa atttcattat taataacgtg atcagcaata aaaggttcca ctacctgatc
     3541 agatacggtg atcctgccca cctccatgag atcgccaaaa acgaggccgt ggtgaagttc
     3601 gtgctcggca ggatcgctga catccagaaa aaacagggcc agaacggcaa gaaccagatc
     3661 gacaggtact acgaaacttg tatcggaaag gataagggca agagcgtgag cgaaaaggtg
     3721 gacgctctca caaagatcat caccggaatg aactacgacc aattcgacaa gaaaaggagc
     3781 gtcattgagg acaccggcag ggaaaacgcc gagagggaga agtttaaaaa gatcatcagc
     3841 ctgtacctca ccgtgatcta ccacatcctc aagaatattg tcaatatcaa cgccaggtac
     3901 gtcatcggat tccattgcgt cgagcgtgat gctcaactgt acaaggagaa aggctacgac
     3961 atcaatctca agaaactgga agagaaggga ttcagctccg tcaccaagct ctgcgctggc
     4021 attgatgaaa ctgcccccga taagagaaag gacgtggaaa aggagatggc tgaaagagcc
     4081 aaggagagca ttgacagcct cgagagcgcc aaccccaagc tgtatgccaa ttacatcaaa
     4141 tacagcgacg agaagaaagc cgaggagttc accaggcaga ttaacaggga gaaggccaaa
     4201 accgccctga acgcctacct gaggaacacc aagtggaatg tgatcatcag ggaggacctc
     4261 ctgagaattg acaacaagac atgtaccctg ttcagaaaca aggccgtcca cctggaagtg
     4321 gccaggtatg tccacgccta tatcaacgac attgccgagg tcaattccta cttccaactg
     4381 taccattaca tcatgcagag aattatcatg aatgagaggt acgagaaaag cagcggaaag
     4441 gtgtccgagt acttcgacgc tgtgaatgac gagaagaagt acaacgatag gctcctgaaa
     4501 ctgctgtgtg tgcctttcgg ctactgtatc cccaggttta agaacctgag catcgaggcc
     4561 ctgttcgata ggaacgaggc cgccaagttc gacaaggaga aaaagaaggt gtccggcaat
     4621 tccggatccg gacctaagaa aaagaggaag gtggcggccg cttacccata cgatgttcca
     4681 gattacgctg ctagcggcTC AGGCgagggc agaggaagtc tgctaacatg cggtgacgtc
     4741 gaggagaatc ctggcccagc gtCCgtgagc aagggcgagg aggataacat ggccatcatc
     4801 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     4861 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     4921 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     4981 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     5041 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     5101 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     5161 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     5221 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     5281 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     5341 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     5401 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     5461 ctgtacaagT AAtgctgagg ctaactgaaa cacggaagga gacaatacag gaaggaacgg
     5521 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5581 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5641 ttatcatgtc tgtcgtctat gatgaggatg ttggtggaga gcatgtggag gaagtggata
     5701 gggaaggttg tagagtagat ccggttgaag tgatgaggat aggaggagga ctgcacatat
     5761 tgatcctcgt cacaaccaca ccagcacacc tcaaatccca caccactccc acaattacca
     5821 ttcactcaac aaactcacac atcccacgat aacgaattca agcttgatat cattcaggac
     5881 gagcctcaga ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg
     5941 ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6001 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6061 gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6121 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6181 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6241 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6301 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6361 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6421 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6481 tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca
     6541 ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc
     6601 aggtgattac atttgggccc tcattaccaa tgcttaatca gtgaggcacc tatctcagcg
     6661 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg
     6721 cgggagggct taccatctgg ccccagtgct g
//