LOCUS       sequence-136524-        3441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector expression sgCentromere-20xPBSc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3441)
  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 3441)
  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: pAC1454_pX-sgCentromere-25xPBSc
FEATURES             Location/Qualifiers
     source          1..3441
                     /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   704..844
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   704..833
                     /label=AAV2 ITR
     rep_origin      919..1374
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1006..1025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1216..1237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1391..1410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1510..1532
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1570..1588)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1656..1760
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1761..1829,1830..2621)
                     /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(1979..1998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2792..3380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3281..3300
                     /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 aaacaccgtt gaggccttcg ttggaaacgt ttaagagcta tgctggaaac
      301 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
      361 gtgccaattg ggtctccaga tttgatgtag ccttgatgta gccttgatgt agccttgatg
      421 tagccttgat gtaagatttg atgtagcctt gatgtagcct tgatgtagcc ttgatgtagc
      481 cttgatgtaa gatttgatgt agccttgatg tagccttgat gtagccttga tgtagccttg
      541 atgtaagatt tgatgtagcc ttgatgtagc cttgatgtag ccttgatgta gccttgatgt
      601 aagatctttt tttgttttag agctagaaat agcaagttaa aataaggcta gtccgtagcg
      661 cgtgcgccaa ttctgcagac aaatggcggc gcgccgcggc cgcaggaacc cctagtgatg
      721 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
      781 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
      841 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
      901 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
      961 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1021 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1081 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     1141 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1201 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1261 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1321 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     1381 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     1441 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     1501 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     1561 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1621 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1681 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1741 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1801 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1861 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1921 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1981 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2041 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2101 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2161 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2221 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2281 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2341 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2401 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2461 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2521 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2581 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2641 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2701 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2761 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2821 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2881 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2941 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3001 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3061 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3121 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3181 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3241 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3301 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3361 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3421 gctggccttt tgctcacatg t
//
