pT7-Cas9

LOCUS       pT7-Cas9                9036 bp DNA     circular SYN 07-MAR-2024
DEFINITION  In vitro transcription vector with the T7 promoter, for producing 
            myc- and FLAG(R)-tagged S. pyogenes Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    pT7-Cas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9036)
  AUTHORS   OriGene
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2024 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1029..5132
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /note="color: #993366"
     CDS             5133..5153
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             5160..5180
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             5202..5231
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             5250..5273
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /note="color: #cc99b2"
                     /translation="DYKDDDDK"
     polyA_signal    5320..5942
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(6091..6679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     polyA_signal    7008..7055
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     CDS             complement(7287..8081)
                     /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))"
                     /note="color: #ccffcc"                     
     promoter        complement(8116..8473)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: LEFT"
     promoter        complement(8475..8579)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      join(8606..9036,1..25)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat ggataagaaa tactcaatag gactggatat tggcacaaat agcgtcggat
     1081 gggctgtgat cactgatgaa tataaggttc cttctaaaaa gttcaaggtt ctgggaaata
     1141 cagaccgcca cagtatcaaa aaaaatctta taggggctct tctgtttgac agtggagaga
     1201 cagccgaagc tactagactc aaacggacag ctaggagaag gtatacaaga cggaagaata
     1261 ggatttgtta tctccaggag attttttcaa atgagatggc caaagtggat gatagtttct
     1321 ttcatagact tgaagagtct tttttggtgg aagaagacaa gaagcatgaa agacatccta
     1381 tttttggaaa tatagtggat gaagttgctt atcacgagaa atatccaact atctatcatc
     1441 tgagaaaaaa attggtggat tctactgata aagccgattt gcgcctgatc tatttggccc
     1501 tggcccacat gattaagttt agaggtcatt ttttgattga gggcgatctg aatcctgata
     1561 atagtgatgt ggacaaactg tttatccagt tggtgcaaac ctacaatcaa ctgtttgaag
     1621 aaaaccctat taacgcaagt ggagtggatg ctaaagccat tctttctgca agattgagta
     1681 aatcaagaag actggaaaat ctcattgctc agctccccgg tgagaagaaa aatggcctgt
     1741 ttgggaatct cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt
     1801 tggcagaaga tgctaaactc cagctttcaa aagatactta cgatgatgat ctggataatc
     1861 tgttggctca aattggagat caatatgctg atttgttttt ggcagctaag aatctgtcag
     1921 atgctattct gctttcagac atcctgagag tgaatactga aataactaag gctcccctgt
     1981 cagcttcaat gattaaacgc tacgatgaac atcatcaaga cttgactctt ctgaaagccc
     2041 tggttagaca acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg
     2101 gatatgcagg ttatattgat ggcggcgcaa gccaagaaga attttataaa tttatcaaac
     2161 caattctgga aaaaatggat ggtactgagg aactgttggt gaaactgaat agagaagatt
     2221 tgctgcgcaa gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg
     2281 agctgcatgc tattttgaga agacaagaag acttttatcc atttctgaaa gacaatagag
     2341 agaagattga aaaaatcttg acttttagga ttccttatta tgttggtcca ttggccagag
     2401 gcaatagtag gtttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt
     2461 ttgaagaagt tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacaaact
     2521 ttgataaaaa tcttccaaat gaaaaagtgc tgccaaaaca tagtttgctt tatgagtatt
     2581 ttaccgttta taacgaattg acaaaggtca aatatgttac tgaaggaatg agaaaaccag
     2641 catttctttc aggtgaacag aagaaagcca ttgttgatct gctcttcaaa acaaatagga
     2701 aagtgaccgt taagcaactg aaagaagatt atttcaaaaa aatagaatgt tttgatagtg
     2761 ttgaaatttc aggagttgaa gatagattta atgcttcact gggtacatac catgatttgc
     2821 tgaaaattat taaagataaa gattttttgg ataatgaaga aaatgaagac atcctggagg
     2881 atattgttct gacattgacc ctgtttgaag atagggagat gattgaggaa agacttaaaa
     2941 catacgctca cctctttgat gataaggtga tgaaacagct taaaagacgc agatatactg
     3001 gttggggaag gttgtccaga aaattgatta atggtattag ggataagcaa tctggcaaaa
     3061 caatactgga ttttttgaaa tcagatggtt ttgccaatcg caattttatg cagctcatcc
     3121 atgatgatag tttgacattt aaagaagaca tccaaaaagc acaagtgtct ggacaaggcg
     3181 atagtctgca tgaacatatt gcaaatctgg ctggtagccc tgctattaaa aaaggtattc
     3241 tccagactgt gaaagttgtt gatgaattgg tcaaagtgat ggggcggcat aagccagaaa
     3301 atatcgttat tgaaatggca agagaaaatc agacaactca aaagggccag aaaaattcca
     3361 gagagaggat gaaaagaatc gaagaaggta tcaaagaact gggaagtcag attcttaaag
     3421 agcatcctgt tgaaaatact caattgcaaa atgaaaagct ctatctctat tatctccaaa
     3481 atggaagaga tatgtatgtg gaccaagaac tggatattaa taggctgagt gattatgatg
     3541 tcgatcacat tgttccacaa agtttcctta aagacgattc aatagacaat aaggtcctga
     3601 ccaggtctga taaaaataga ggtaaatccg ataacgttcc aagtgaagaa gtggtcaaaa
     3661 agatgaaaaa ctattggaga caacttctga acgccaagct gatcactcaa aggaagtttg
     3721 ataatctgac caaagctgaa agaggaggtt tgagtgaact tgataaagct ggttttatca
     3781 aacgccaatt ggttgaaact cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc
     3841 gcatgaatac taaatacgat gaaaatgata aacttattag agaggttaaa gtgattaccc
     3901 tgaaatctaa actggtttct gacttcagaa aagatttcca attctataaa gtgagagaga
     3961 ttaacaatta ccatcatgcc catgatgcct atctgaatgc cgtcgttgga actgctttga
     4021 ttaagaaata tccaaaactt gaaagcgagt ttgtctatgg tgattataaa gtttatgatg
     4081 ttaggaaaat gattgctaag tctgagcaag aaataggcaa agcaaccgca aagtatttct
     4141 tttactctaa tatcatgaac ttcttcaaaa cagaaattac acttgcaaat ggagagattc
     4201 gcaaacgccc tctgatcgaa actaatgggg aaactggaga aattgtctgg gataaaggga
     4261 gagattttgc cacagtgcgc aaagtgttgt ccatgcccca agtcaatatc gtcaagaaaa
     4321 cagaagtgca gacaggcgga ttctctaagg agtcaattct gccaaaaaga aattccgaca
     4381 agctgattgc taggaaaaaa gactgggacc caaaaaaata tggtggtttt gatagtccaa
     4441 ccgtggctta ttcagtcctg gtggttgcta aggtggaaaa agggaaatcc aagaagctga
     4501 aatccgttaa agagctgctg gggatcacaa ttatggaaag aagttccttt gaaaaaaatc
     4561 ccattgactt tctggaagct aaaggatata aggaagttaa aaaagacctg atcattaaac
     4621 tgcctaaata tagtcttttt gagctggaaa acggtaggaa acggatgctg gctagtgccg
     4681 gagaactgca aaaaggaaat gagctggctc tgccaagcaa atatgtgaat tttctgtatc
     4741 tggctagtca ttatgaaaag ttgaagggta gtccagaaga taacgaacaa aaacaattgt
     4801 ttgtggagca gcataagcat tatctggatg agattattga gcaaatcagt gaattttcta
     4861 agagagttat tctggcagat gccaatctgg ataaagttct tagtgcatat aacaaacata
     4921 gagacaaacc aataagagaa caagcagaaa atatcattca tctgtttacc ttgaccaatc
     4981 ttggagcacc cgctgctttt aaatactttg atacaacaat tgataggaaa agatatacct
     5041 ctacaaaaga agttctggat gccactctta tccatcaatc catcactggt ctttatgaaa
     5101 cacgcattga tttgagtcag ctgggaggtg accccaagaa aaaacgcaag gtggaagatc
     5161 ctaagaaaaa gcggaaagtg gacacgcgta cgcggccgct cgagcagaaa ctcatctcag
     5221 aagaggatct ggcagcaaat gatatcctgg attacaagga tgacgacgat aaggtttaaa
     5281 cggccggccg cggtcatagc tgtttcctga acagatcccg ggtggcatcc ctgtgacccc
     5341 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     5401 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     5461 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     5521 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     5581 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     5641 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     5701 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     5761 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag
     5821 caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg
     5881 agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa
     5941 ttgggtacgc ggccagcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6001 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6061 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6121 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6181 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6241 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6301 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6361 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6421 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6481 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6541 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6601 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga
     6841 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     6901 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg
     6961 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     7021 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     7081 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag
     7141 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     7201 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     7261 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     7321 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     7381 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcgatccgcc
     7441 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     7501 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg
     7561 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     7621 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     7681 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     7741 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     7801 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     7861 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg
     7921 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     7981 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     8041 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     8101 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     8161 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     8221 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     8281 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     8341 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     8401 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     8461 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8521 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8581 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     8641 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     8701 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     8761 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     8821 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     8881 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     8941 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     9001 aatgagctga tttaacaaaa atttaacgcg aatttt
//