LOCUS       sequence-171565-        3006 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses S. Pyogenes guide with PP7 aptamer in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3006)
  AUTHORS   Maji B, Moore CL, Zetsche B, Volz SE, Zhang F, Shoulders MD, 
            Choudhary A
  TITLE     Multidimensional chemical control of CRISPR-Cas9.
  JOURNAL   Nat Chem Biol. 2017 Jan;13(1):9-11. doi: 10.1038/nchembio.2224. Epub
            2016 Oct 31.
  PUBMED    27820801
REFERENCE   2  (bases 1 to 3006)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nchembio.2224"; journalName: "Nat Chem Biol"; date: 
            "2017-01"; volume: "13"; issue: "1"; pages: "9-11"
COMMENT     Sequence Label: PP7 guide
FEATURES             Location/Qualifiers
     source          1..3006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..76)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     267..288
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(442..461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     561..583
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(621..639)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        707..811
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(812..880,881..1672)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1030..1049)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1843..2431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2332..2351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2494..2742
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2494..2514
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2665..2684
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     rep_origin      join(2976..3006,1..425)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
       61 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
      121 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
      181 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
      241 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
      301 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
      361 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
      421 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
      481 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
      541 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
      601 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
      661 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
      721 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
      781 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
      841 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
      901 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
      961 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1021 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1081 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1141 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1201 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1261 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1321 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1381 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1441 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1501 tgataaatct ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga
     1561 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     1621 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     1681 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1741 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1801 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1981 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2461 cctggccttt tgctggcctt ttgctcagct agcgagggcc tatttcccat gattccttca
     2521 tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac
     2581 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
     2641 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
     2701 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgggtcttc gagaagacct
     2761 gttttagagc taggccggag cagacgatat ggcgtcgctc cggcctagca agttaaaata
     2821 aggctagtcc gttatcaact tggccggagc agacgatatg gcgtcgctcc ggccaagtgg
     2881 caccgagtcg gtgctttttt tgaattctga tgcggtattt tctccttacg catctgtgcg
     2941 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3001 cgcggc
//
