LOCUS       sequence-437335-        2368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pegRNA 1 plasmid used for PASSIGE-mediated Bxb1 attP installation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2368)
  AUTHORS   Pandey S, Gao XD, Krasnow NA, McElroy A, Tao YA, Duby JE, Steinbeck 
            BJ, McCreary J, Pierce SE, Tolar J, Meissner TB, Chaikof EL, Osborn 
            MJ, Liu DR
  TITLE     Efficient site-specific integration of large genes in mammalian 
            cells via continuously evolved recombinases and prime editing.
  JOURNAL   Nat Biomed Eng. 2024 Jun 10. doi: 10.1038/s41551-024-01227-1.
  PUBMED    38858586
REFERENCE   2  (bases 1 to 2368)
  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 Biomed 
            Eng. 2024 Jun 10. doi: 10.1038/s41551-024-01227-1."
COMMENT     Sequence Label: AAVS1 pegRNA1 with trimmed attP
FEATURES             Location/Qualifiers
     source          1..2368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     340..359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_RNA        complement(632..707)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(728..976)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(786..805)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(956..976)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        1083..1187
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1188..1256,1257..2048)
                     /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(1406..1425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2219..2368,1..439)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
       61 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      121 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      181 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      241 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      301 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      361 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      421 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      481 ctggcctttt gctcacatgt catatgtaga gaggtacctc gagcggccca agcttaaaaa
      541 aattctagtt ggtttaacgc gtaactagat agaaccgcgg ccccccaccg ccaggtttgt
      601 ctggtcaacc accgcggtct cagtggtgta cgcaccgact cggtgccact ttttcaagtt
      661 gataacggac tagccttatt ttaacttgct atttctagct ctaaaactgg ggcggtgggg
      721 ggccagcggt gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc
      781 aagttacggt aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc
      841 caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca
      901 aattaattct aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca
      961 tgggaaatag gccctcttcc tgcccgacct tggtaccgga tccagtcgac tgaattggtt
     1021 cctttaaagc ctgctttttt gtacagctag cgacgtcagg tggcactttt cggggaaatg
     1081 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1141 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1201 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1261 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1321 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1381 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1441 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1501 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1561 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1621 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1681 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1741 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1801 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1861 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1921 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1981 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2041 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2101 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2161 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2221 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2281 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2341 gcagagcgca gataccaaat actgttct
//
