pCas-Scramble

LOCUS       pCas-Scramble           7997 bp DNA     circular SYN 03-NOV-2024
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7997)
  AUTHORS   Ming
  TITLE     Direct Submission
  JOURNAL   Exported Nov 3, 2024 from SnapGene 6.0.2 to Vector NTI(R) format
            https://www.biofeng.com
COMMENT     VNTDATE|1073001600|
COMMENT     VNTDBDATE|1073001600|
COMMENT     VNTNAME|pCas-Scramble|
COMMENT     VNTAUTHORNAME|Ming|
FEATURES             Location/Qualifiers
     promoter        90..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        359..434
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        486..865
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        866..1069
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        1095..1113
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1172..5275
                     /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"
     CDS             5276..5296
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
     CDS             5303..5323
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
     CDS             5345..5374
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
     CDS             5393..5416
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
     primer_bind     complement(5436..5452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5463..6085
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     rep_origin      complement(6206..6794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6958..7818)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     primer_bind     7933..7949
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7954..7972
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaattcccca gtggaaagac gcgcaggcaa aacgcaccac gtgacggagc gtgaccgcgc
       61 gccgagcgcg cgccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt
      121 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa
      181 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
      241 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
      301 tcttgggttt atatatcttg tggaaaggac gcgggatcgc actaccagag ctaactcagt
      361 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
      421 caccgagtcg gtgctttttt tggtgtacat ttatattggc tcatgtccaa tatgaccgcc
      481 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      541 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      601 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      661 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      721 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
      781 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
      841 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
      901 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      961 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     1021 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     1081 ccgtcagaat tttgtaatac gactcactat agggcggccg ggaattcgtc gactggaacc
     1141 ggtaccgagg agatctgccg ccgcgatcgc catggataag aaatactcaa taggactgga
     1201 tattggcaca aatagcgtcg gatgggctgt gatcactgat gaatataagg ttccttctaa
     1261 aaagttcaag gttctgggaa atacagaccg ccacagtatc aaaaaaaatc ttataggggc
     1321 tcttctgttt gacagtggag agacagccga agctactaga ctcaaacgga cagctaggag
     1381 aaggtataca agacggaaga ataggatttg ttatctccag gagatttttt caaatgagat
     1441 ggccaaagtg gatgatagtt tctttcatag acttgaagag tcttttttgg tggaagaaga
     1501 caagaagcat gaaagacatc ctatttttgg aaatatagtg gatgaagttg cttatcacga
     1561 gaaatatcca actatctatc atctgagaaa aaaattggtg gattctactg ataaagccga
     1621 tttgcgcctg atctatttgg ccctggccca catgattaag tttagaggtc attttttgat
     1681 tgagggcgat ctgaatcctg ataatagtga tgtggacaaa ctgtttatcc agttggtgca
     1741 aacctacaat caactgtttg aagaaaaccc tattaacgca agtggagtgg atgctaaagc
     1801 cattctttct gcaagattga gtaaatcaag aagactggaa aatctcattg ctcagctccc
     1861 cggtgagaag aaaaatggcc tgtttgggaa tctcattgct ttgtcattgg gtttgacccc
     1921 taattttaaa tcaaattttg atttggcaga agatgctaaa ctccagcttt caaaagatac
     1981 ttacgatgat gatctggata atctgttggc tcaaattgga gatcaatatg ctgatttgtt
     2041 tttggcagct aagaatctgt cagatgctat tctgctttca gacatcctga gagtgaatac
     2101 tgaaataact aaggctcccc tgtcagcttc aatgattaaa cgctacgatg aacatcatca
     2161 agacttgact cttctgaaag ccctggttag acaacaactt ccagaaaagt ataaagaaat
     2221 cttttttgat caatcaaaaa acggatatgc aggttatatt gatggcggcg caagccaaga
     2281 agaattttat aaatttatca aaccaattct ggaaaaaatg gatggtactg aggaactgtt
     2341 ggtgaaactg aatagagaag atttgctgcg caagcaacgg acctttgaca acggctctat
     2401 tccccatcaa attcacttgg gtgagctgca tgctattttg agaagacaag aagactttta
     2461 tccatttctg aaagacaata gagagaagat tgaaaaaatc ttgactttta ggattcctta
     2521 ttatgttggt ccattggcca gaggcaatag taggtttgca tggatgactc ggaagtctga
     2581 agaaacaatt accccatgga attttgaaga agttgtcgat aaaggtgctt cagctcaatc
     2641 atttattgaa cgcatgacaa actttgataa aaatcttcca aatgaaaaag tgctgccaaa
     2701 acatagtttg ctttatgagt attttaccgt ttataacgaa ttgacaaagg tcaaatatgt
     2761 tactgaagga atgagaaaac cagcatttct ttcaggtgaa cagaagaaag ccattgttga
     2821 tctgctcttc aaaacaaata ggaaagtgac cgttaagcaa ctgaaagaag attatttcaa
     2881 aaaaatagaa tgttttgata gtgttgaaat ttcaggagtt gaagatagat ttaatgcttc
     2941 actgggtaca taccatgatt tgctgaaaat tattaaagat aaagattttt tggataatga
     3001 agaaaatgaa gacatcctgg aggatattgt tctgacattg accctgtttg aagataggga
     3061 gatgattgag gaaagactta aaacatacgc tcacctcttt gatgataagg tgatgaaaca
     3121 gcttaaaaga cgcagatata ctggttgggg aaggttgtcc agaaaattga ttaatggtat
     3181 tagggataag caatctggca aaacaatact ggattttttg aaatcagatg gttttgccaa
     3241 tcgcaatttt atgcagctca tccatgatga tagtttgaca tttaaagaag acatccaaaa
     3301 agcacaagtg tctggacaag gcgatagtct gcatgaacat attgcaaatc tggctggtag
     3361 ccctgctatt aaaaaaggta ttctccagac tgtgaaagtt gttgatgaat tggtcaaagt
     3421 gatggggcgg cataagccag aaaatatcgt tattgaaatg gcaagagaaa atcagacaac
     3481 tcaaaagggc cagaaaaatt ccagagagag gatgaaaaga atcgaagaag gtatcaaaga
     3541 actgggaagt cagattctta aagagcatcc tgttgaaaat actcaattgc aaaatgaaaa
     3601 gctctatctc tattatctcc aaaatggaag agatatgtat gtggaccaag aactggatat
     3661 taataggctg agtgattatg atgtcgatca cattgttcca caaagtttcc ttaaagacga
     3721 ttcaatagac aataaggtcc tgaccaggtc tgataaaaat agaggtaaat ccgataacgt
     3781 tccaagtgaa gaagtggtca aaaagatgaa aaactattgg agacaacttc tgaacgccaa
     3841 gctgatcact caaaggaagt ttgataatct gaccaaagct gaaagaggag gtttgagtga
     3901 acttgataaa gctggtttta tcaaacgcca attggttgaa actcgccaaa tcactaagca
     3961 tgtggcacaa attttggata gtcgcatgaa tactaaatac gatgaaaatg ataaacttat
     4021 tagagaggtt aaagtgatta ccctgaaatc taaactggtt tctgacttca gaaaagattt
     4081 ccaattctat aaagtgagag agattaacaa ttaccatcat gcccatgatg cctatctgaa
     4141 tgccgtcgtt ggaactgctt tgattaagaa atatccaaaa cttgaaagcg agtttgtcta
     4201 tggtgattat aaagtttatg atgttaggaa aatgattgct aagtctgagc aagaaatagg
     4261 caaagcaacc gcaaagtatt tcttttactc taatatcatg aacttcttca aaacagaaat
     4321 tacacttgca aatggagaga ttcgcaaacg ccctctgatc gaaactaatg gggaaactgg
     4381 agaaattgtc tgggataaag ggagagattt tgccacagtg cgcaaagtgt tgtccatgcc
     4441 ccaagtcaat atcgtcaaga aaacagaagt gcagacaggc ggattctcta aggagtcaat
     4501 tctgccaaaa agaaattccg acaagctgat tgctaggaaa aaagactggg acccaaaaaa
     4561 atatggtggt tttgatagtc caaccgtggc ttattcagtc ctggtggttg ctaaggtgga
     4621 aaaagggaaa tccaagaagc tgaaatccgt taaagagctg ctggggatca caattatgga
     4681 aagaagttcc tttgaaaaaa atcccattga ctttctggaa gctaaaggat ataaggaagt
     4741 taaaaaagac ctgatcatta aactgcctaa atatagtctt tttgagctgg aaaacggtag
     4801 gaaacggatg ctggctagtg ccggagaact gcaaaaagga aatgagctgg ctctgccaag
     4861 caaatatgtg aattttctgt atctggctag tcattatgaa aagttgaagg gtagtccaga
     4921 agataacgaa caaaaacaat tgtttgtgga gcagcataag cattatctgg atgagattat
     4981 tgagcaaatc agtgaatttt ctaagagagt tattctggca gatgccaatc tggataaagt
     5041 tcttagtgca tataacaaac atagagacaa accaataaga gaacaagcag aaaatatcat
     5101 tcatctgttt accttgacca atcttggagc acccgctgct tttaaatact ttgatacaac
     5161 aattgatagg aaaagatata cctctacaaa agaagttctg gatgccactc ttatccatca
     5221 atccatcact ggtctttatg aaacacgcat tgatttgagt cagctgggag gtgaccccaa
     5281 gaaaaaacgc aaggtggaag atcctaagaa aaagcggaaa gtggacacgc gtacgcggcc
     5341 gctcgagcag aaactcatct cagaagagga tctggcagca aatgatatcc tggattacaa
     5401 ggatgacgac gataaggttt aaacggccgg ccgcggtcat agctgtttcc tgaacagatc
     5461 ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     5521 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     5581 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     5641 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     5701 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     5761 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagaggc
     5821 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     5881 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     5941 tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc
     6001 atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt
     6061 ccactcagta gatgcctgtt gaattgggta cgcggccagc ggcgagcggt atcagctcac
     6121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtcc
     6181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6841 acgcgtaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6901 atccttttgc ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7861 ttgtctcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacacg
     7921 ggccagagct gccaggaaac agctatgacc atgtaatacg actcactata ggggatatca
     7981 gctggatggc agttaac
//