LOCUS       sequence-367469-        2738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control vector for coselection for prime editing in human cells. 
            Tandem expression of ATP1A1 Q118R-G4 pegRNA and EBFP_To_EGFP epegRNA
            (tevopreq1) from two independent U6 promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2738)
  AUTHORS   Levesque S, Mayorga D, Fiset JP, Goupil C, Duringer A, Loiselle A, 
            Bouchard E, Agudelo D, Doyon Y
  TITLE     Marker-free co-selection for successive rounds of prime editing in 
            human cells.
  JOURNAL   Nat Commun. 2022 Oct 7;13(1):5909. doi: 10.1038/s41467-022-33669-z.
  PUBMED    36207338
REFERENCE   2  (bases 1 to 2738)
  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"; journalName: "Nat 
            Commun."; date: "2022-10-7"; volume: "13(1):5909. doi"; pages: " 
            10.1038/s41467-022-33669-z"
COMMENT     Sequence Label: 
            ATP1A1_G4_Q118R_Std_BFP-to-EGFP_Dual_epegRNA_tevopreQ1
FEATURES             Location/Qualifiers
     source          1..2738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..320
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        341..416
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(582..1170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(662..681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1341..2132,2133..2201))
                     /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     1964..1983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2202..2306)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2369..2617
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        2638..2713
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tttttttaag cttgggccgc tcgaggtacc tctctacgtc gactggatcc ggtaccaagg
       61 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
      121 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
      181 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      241 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      301 tgtggaaagg acgaaacacc ggctgaagca ctgcacgccg gttttagagc tagaaatagc
      361 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgcccct
      421 gacctacggc gtgcagtgct tcgcggttct atctagttac gcgttaaacc aactagaatt
      481 tttttaagct tgggccgctc gaggtacctc tctacatatg acatgtgagc aaaaggccag
      541 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      601 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      661 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      721 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      781 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      841 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      901 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      961 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1021 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1081 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1141 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1201 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1261 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1321 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1381 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1441 gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct
     1501 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1561 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1621 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1681 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1741 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1801 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1861 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1921 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1981 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2041 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2101 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2161 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2221 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2281 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgct
     2341 agctgtacat accaaggtcg ggcaggaaga gggcctattt cccatgattc cttcatattt
     2401 gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa
     2461 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     2521 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     2581 cttggcttta tatatcttgt ggaaaggacg aaacaccgtt cctcttctgt agcagctgtt
     2641 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     2701 accgagtcgg tgctagcatc agagctgcta cagaagag
//
