LOCUS       sequence-134361-        3382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector for expression of sgRNA-15xPBSw.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3382)
  AUTHORS   Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, 
            Taghbalout A, Wang H
  TITLE     Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation 
            and genomic labeling.
  JOURNAL   Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 
            Jan 15.
  PUBMED    26768771
REFERENCE   2  (bases 1 to 3382)
  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"; doi: "10.1038/cr.2016.3";
            journalName: "Cell Res"; date: "2016-02"; volume: "26"; issue: "2"; 
            pages: "254-7"
COMMENT     Sequence Label: pAC1428-pX-sgRNA-15xPBSw
FEATURES             Location/Qualifiers
     source          1..3382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..257
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9..29
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     180..199
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   645..785
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   645..774
                     /label=AAV2 ITR
     rep_origin      860..1315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(947..966)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1157..1178
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1332..1351)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1451..1473
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1511..1529)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1597..1701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1702..1770,1771..2562)
                     /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(1920..1939)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2733..3321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3222..3241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcgcgccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
       61 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
      121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
      181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
      241 ggaaaggacg aaacaccggg tcttcgagaa gacctgttta agagctatgc tggaaacagc
      301 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      361 ccaattgggt ctccagattg tatatagcct gtatatagcc tgtatatagc ctgtatatag
      421 cctgtatata agattgtata tagcctgtat atagcctgta tatagcctgt atatagcctg
      481 tatataagat tgtatatagc ctgtatatag cctgtatata gcctgtatat agcctgtata
      541 taagatcttt ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtagc
      601 gcgtgcgcca attctgcaga caaatggcgg cgcgccgcgg ccgcaggaac ccctagtgat
      661 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
      721 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
      781 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      841 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      901 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      961 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1021 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     1081 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1141 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1201 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1261 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     1321 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     1381 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     1441 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     1501 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1561 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1621 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1681 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1741 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1801 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1861 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1921 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1981 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2041 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2101 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2161 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2221 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2281 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2341 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2401 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     2461 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2521 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2581 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2641 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2701 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2761 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2821 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3241 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3301 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3361 tgctggcctt ttgctcacat gt
//
