LOCUS       sequence-134363-        3739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector for expression of sgRNA-47xPBSb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3739)
  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 3739)
  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: pAC1426-pX-sgRNA-47xPBSb
FEATURES             Location/Qualifiers
     source          1..3739
                     /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   1002..1142
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1002..1131
                     /label=AAV2 ITR
     rep_origin      1217..1672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1304..1323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1514..1535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1689..1708)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1808..1830
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1868..1886)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1954..2058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2059..2127,2128..2919)
                     /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(2277..2296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3090..3678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3579..3598
                     /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 ctccagattt gatatagcct tgatatagcc ttgatatagc cttgatatag
      421 ccttgatata agatttgata tagccttgat atagccttga tatagccttg atatagcctt
      481 gatataagat ttgatatacc ttgatatagc cttgatatag ccttgatata gccttgatat
      541 aagatttgat atagccttga tatagccttg atatagcctt gatatagcct tgatatagcc
      601 ttgatatagc cttgatataa gatttgatat agccttgata tagccttgat atagccttga
      661 tatagccttg atataagatt tgatatagcc ttgatatagc cttgatatag ccttgatata
      721 gccttgatat aagatttgat atagccttga tatagccttg atatagcctt gatatagcct
      781 tgatataaga tttgatatag ccttgatata gccttgatat agccttgata tagccttgat
      841 ataagatttg atatagcctt gatatagcct tgatatagcc ttgatatagc cttgatataa
      901 gatctttttt tgttttagag ctagaaatag caagttaaaa taaggctagt ccgtagcgcg
      961 tgcgccaatt ctgcagacaa atggcggcgc gccgcggccg caggaacccc tagtgatgga
     1021 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     1081 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     1141 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     1201 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1261 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1321 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1381 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     1441 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1501 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     1561 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1621 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     1681 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     1741 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1801 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     1861 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1921 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1981 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2041 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2101 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2161 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2221 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2281 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     2341 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2401 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2461 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2521 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2581 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2641 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2701 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2761 gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2821 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2881 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2941 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3001 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3061 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3121 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3181 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3241 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3301 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3361 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3421 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3481 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3541 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3601 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3661 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3721 tggccttttg ctcacatgt
//
