LOCUS       sequence-351758-        2377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ACTB N-term PBS 13 RT 29 Bxb1 AttB 46 atgRNA with v2 scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2377)
  AUTHORS   Yarnall MTN, Ioannidi EI, Schmitt-Ulms C, Krajeski RN, Lim J, 
            Villiger L, Zhou W, Jiang K, Garushyants SK, Roberts N, Zhang L, 
            Vakulskas CA, Walker JA 2nd, Kadina AP, Zepeda AE, Holden K, Ma H, 
            Xie J, Gao G, Foquet L, Bial G, Donnelly SK, Miyata Y, Radiloff DR, 
            Henderson JM, Ujita A, Abudayyeh OO, Gootenberg JS
  TITLE     Drag-and-drop genome insertion of large sequences without 
            double-strand DNA cleavage using CRISPR-directed integrases.
  JOURNAL   Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4.
  PUBMED    36424489
REFERENCE   2  (bases 1 to 2377)
  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 
            Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4."
COMMENT     Sequence Label: pDY0663 ACTB atgRNA with v2 scaffold
FEATURES             Location/Qualifiers
     source          1..2377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(38..626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(118..137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(797..1588,1589..1657))
                     /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     1420..1439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1658..1762)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1869..2117
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1869..1889
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2040..2059
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    2257..2294
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
ORIGIN
        1 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
       61 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      421 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
      481 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      541 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      721 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      901 gcttaccatc tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag
      961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1381 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgctagct
     1801 gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac tggatccggt accaaggtcg
     1861 ggcaggaaga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     1921 ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     1981 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     2041 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     2101 ggaaaggacg aaacaccgct attctcgcag ctcaccagtt tgagagctat gctggaaaca
     2161 gcatagcaag ttcaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     2221 tgcgacgagc gcggcgatat catcatccat ggccggatga tcctgacgac ggagaccgcc
     2281 gtcgtcgaca agccggcctg agctgcgaga atttttttaa gcttgggccg ctcgaggtac
     2341 ctctctacat atgacatgtg agcaaaaggc cagcaaa
//
