LOCUS       sequence-155834-        5067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid that expresses Cas9C in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5067)
  AUTHORS   Chew WL, Tabebordbar M, Cheng JK, Mali P, Wu EY, Ng AH, Zhu K, 
            Wagers AJ, Church GM
  TITLE     A multifunctional AAV-CRISPR-Cas9 and its host response.
  JOURNAL   Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993.
  PUBMED    27595405
REFERENCE   2  (bases 1 to 5067)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993."
COMMENT     Sequence Label: pSMVP-Cas9C
FEATURES             Location/Qualifiers
     source          1..5067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..59)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     enhancer        289..668
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        669..872
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     823..843
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          964..1096
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1126..1135
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1130..1139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1736..1747
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3265..3285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3296..3315)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3325..3406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3350..3369
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3604..4192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3684..3703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4273..5067)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4273..5067)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4384..4403)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4994..5013
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 aatattattg aagcatttat cagggttcgt ctcgctgtgg aatgtgtgtc agttagggtg
       61 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      121 agcaaccagt cccggtctcc tcccatgcat gtcaatattg gccattagcc atattattca
      181 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
      241 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat
      301 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      361 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      421 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      481 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      541 tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      601 agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      661 ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac
      721 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      781 aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc
      841 gtgtacggtg ggaggtctat ataagcagag gtcgtttagt gaaccgtcag atcactagta
      901 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct cgactgatca
      961 caggtaagta tcaaggttac aagacaggtt taaggaggcc aatagaaact gggcttgtcg
     1021 agacagagaa gattcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     1081 gcctttctct ccacaggggt accgaagccg ctagcgctac cggtcgccac catggcggcg
     1141 gcgtgcccgg aactgcgtca gctggcgcag agcgatgtgt attgggatcc gattgtgagc
     1201 attgaaccgg atggcgtgga agaagtgttt gatctgaccg tgccgggccc gcataacttt
     1261 gtggcgaacg atattattgc gcataactct ggccaggggg acagtcttca cgagcacatc
     1321 gctaatcttg caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg
     1381 gatgaactcg tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc
     1441 cgagagaacc aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt
     1501 gaagagggta taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc
     1561 cagcttcaga atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg
     1621 gatcaggaac tggacatcaa tcggctctcc gactacgacg tggatcatat cgtgccccag
     1681 tcttttctca aagatgattc tattgataat aaagtgttga caagatccga taaaaataga
     1741 gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg
     1801 cagctgctga acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa
     1861 cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca
     1921 cgccagatca ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat
     1981 gaaaatgaca aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtctca
     2041 gatttcagaa aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg
     2101 catgatgcct acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctt
     2161 gaatctgaat ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag
     2221 tctgagcagg aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat
     2281 tttttcaaga ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa
     2341 acaaacggag aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgg
     2401 aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc
     2461 ttctccaagg aaagtatcct cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa
     2521 gattgggacc ccaagaaata cggcggattc gattctccta cagtcgctta cagtgtactg
     2581 gttgtggcca aagtggagaa agggaagtct aaaaaactca aaagcgtcaa ggaactgctg
     2641 ggcatcacaa tcatggagcg atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg
     2701 aaaggatata aagaggtcaa aaaagacctc atcattaagc ttcccaagta ctctctcttt
     2761 gagcttgaaa acggccggaa acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac
     2821 gagctggcac tgccctctaa atacgttaat ttcttgtatc tggccagcca ctatgaaaag
     2881 ctcaaagggt ctcccgaaga taatgagcag aagcagctgt tcgtggaaca acacaaacac
     2941 taccttgatg agatcatcga gcaaataagc gaattctcca aaagagtgat cctcgccgac
     3001 gctaacctcg ataaggtgct ttctgcttac aataagcaca gggataagcc catcagggag
     3061 caggcagaaa acattatcca cttgtttact ctgaccaact tgggcgcgcc tgcagccttc
     3121 aagtacttcg acaccaccat agacagaaag cggtacacct ctacaaagga ggtcctggac
     3181 gccacactga ttcatcagtc aattacgggg ctctatgaaa caagaatcga cctctctcag
     3241 ctcggtggag acagcagggc tgaccccaag aagaagagga aggtgtctcg agctcgatga
     3301 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3361 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3421 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3481 cctctacaaa tgtggtactt aagaggggga gaccaaaggg cgagacgtta aggcctcacg
     3541 tgacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3721 gcgctctcct gttccgaccc tgccgcttac gggatacctg tccgcctttc tcccttcggg
     3781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4021 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4261 ggtcatgccg tctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     4321 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     4381 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     4441 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     4501 gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag
     4561 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     4621 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     4681 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     4741 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     4801 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     4861 ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt
     4921 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     4981 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     5041 tgcgtgcaat ccatcttgtt caatcat
//
