pU6-BAG3-PegRNA

LOCUS      pU6-BAG3-PegRNA          2307 bp DNA     circular SYN 13-MAR-2026
DEFINITION  Prime editing in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2307)
  TITLE     Liu Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2307)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
FEATURES             Location/Qualifiers
     source          1..2307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     234..266
                     /label=OL209
     primer_bind     complement(234..266)
                     /label=OL300
     misc_feature    251..270
                     /label=BAG3 spacer
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    347..358
                     /label=RTT
     misc_feature    359..373
                     /label=PBS
     primer_bind     complement(360..373)
                     /label=OL209
     rep_origin      complement(477..1065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(557..576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1236..2096)
                     /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     1859..1878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2097..2201)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg actcagccag ataaacagtg gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgcacct gtccacgctg
      361 tttatctggc tgattttttt aagcttgggc cgctcgaggt acctctctac atatgacatg
      421 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      481 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      541 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      601 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      661 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      721 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      781 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      841 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      901 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      961 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1021 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1081 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1141 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1201 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1261 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1321 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagat
     1381 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1441 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1501 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1561 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1621 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1681 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1741 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1801 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1861 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1921 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1981 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2041 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2101 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2161 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2221 ccacctgacg tcgctagctg tacaaaaaag caggctttaa aggaaccaat tcagtcgact
     2281 ggatccggta ccaaggtcgg gcaggaa
//