LOCUS       sequence-367786-        3774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockin of truncated NGFR to target gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3774)
  AUTHORS   Shy BR, Vykunta VS, Ha A, Talbot A, Roth TL, Nguyen DN, Pfeifer WG, 
            Chen YY, Blaeschke F, Shifrut E, Vedova S, Mamedov MR, Chung JJ, Li 
            H, Yu R, Wu D, Wolf J, Martin TG, Castro CE, Ye L, Esensten JH, 
            Eyquem J, Marson A
  TITLE     High-yield genome engineering in primary cells using a hybrid ssDNA 
            repair template and small-molecule cocktails.
  JOURNAL   Nat Biotechnol. 2022 Aug 25:10.1038/s41587-022-01418-8. doi: 
            10.1038/s41587-022-01418-8.
  PUBMED    36008610
REFERENCE   2  (bases 1 to 3774)
  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 Aug 25:10.1038/s41587-022-01418-8. doi: 
            10.1038/s41587-022-01418-8."
COMMENT     Sequence Label: pIG-702pUC19-tNGFR-P2A-LRBA
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1186..1242
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(1620..1636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1620..1636)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1633..1655)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1808..2396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1888..1907)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2513..2537)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2637..3428,3429..3497))
                     /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     3260..3279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3498..3602)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3701..3723
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3715..3732
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3716..3732
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgcctgacct tcgagctcgg tacccgggga acaataacta tttccctctt gtcccctcaa
       61 aaccctaaaa caaaacctag cctatttaac atatatttaa tcttccaata gggtttggcg
      121 ttgttgtcag cctcggggag agagattgga caaatattct ccaagaggag gagggcgacg
      181 ccaaggactt tccacatcaa ctgctttggg gtatctccac aagttggaag agggaccctt
      241 tcgttttgca ttgcgtgtgt tgtgctcatt accagtgcag cgactgccgt cccagggtga
      301 ctctgagttg tcgtttatcg tcagctaacc atgggtgctg gtgcaacggg gagggctatg
      361 gatggaccaa ggctccttct tcttttgctc ttgggcgtta gtctgggtgg agctaaagag
      421 gcatgcccca ccggacttta cacacattcc ggtgagtgtt gcaaggcctg caacctggga
      481 gaaggcgtcg ctcaaccctg cggggcgaat caaaccgtat gcgaaccttg cctcgattct
      541 gttacattct ccgatgtcgt ctctgcgacg gaaccctgca agccatgcac ggaatgtgtc
      601 gggttgcaaa gtatgtcagc tccttgcgtt gaagcagacg atgcggtgtg tagatgtgcc
      661 tatgggtatt atcaagacga aacgacggga cggtgtgaag catgtcgagt atgcgaggcg
      721 ggcagtggcc tcgtcttctc ttgtcaggac aagcaaaaca ctgtctgtga agaatgccct
      781 gatgggactt atagtgatga ggcgaatcac gtcgatccat gtcttccctg tactgtctgt
      841 gaagacactg agagacaatt gcgagagtgt acgcgatggg ccgacgcaga gtgcgaggag
      901 atcccaggtc gatggataac caggtcaacc cctcctgagg gatctgatag taccgcgccc
      961 tccacccaag aacccgaagc tccacctgag caagacctca tagcgtccac agtagcagga
     1021 gttgtcacaa ccgtcatggg aagttcacaa ccagtagtta ccaggggtac cacggataac
     1081 ctcattccgg tgtactgcag cattctcgcc gctgtggttg tcggacttgt tgcatacatt
     1141 gccttcaaac gctggaatag ccgcgccaag cgctcgggtt cgggtgccac caacttcagc
     1201 ctgctgaagc aggccggcga cgtggaggag aaccccggcc ccgctagcga agacaatcgt
     1261 gtcccttctc cccctcctac aggtgatgac gggggaggtg gagggagaga agaaacccct
     1321 actgaagggg gtgcattgtc tctgaaacca gggctcccca tcaggggcat cagaatgaaa
     1381 tttgccgtgt tgaccggttt ggttgaagtt ggagaagtat ccaataggga tattgtagaa
     1441 actgtcttta acctggtaag tccatgaata ttttataagc atatactctt ttcagtgggt
     1501 tggggaacaa gatcttaaag ctggagccat gtggtttact catcctctag agtcgacctg
     1561 cagaggctag gtggaggctc agtgatgata agtctgcgat ggtggatgca tgtgtcatgg
     1621 tcatagctgt ttcctgtgtg aaattgttat ccgctcagag ggcacaatcc tattccgcgc
     1681 tatccgacaa tctccaagac attaggtgga gttcagttcg gcgtatggca tatgtcgctg
     1741 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1801 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1861 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1921 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1981 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2041 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2101 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2161 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2221 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2281 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2341 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2401 atcctttgat cttttctacg gggtctgacg ctctattcaa caaagccgcc gtcccgtcaa
     2461 gtcagcgtaa atgggtaggg ggcttcaaat cgtcctcgtg ataccaattc ggagcctgct
     2521 tttttgtaca aacttgttga taatggcaat tcaaggatct tcacctagat ccttttaaat
     2581 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2641 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2701 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2761 gctgcaatga taccgcgaga gccacgctca ccggctccag atttatcagc aataaaccag
     2821 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2881 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2941 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3001 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3061 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3601 cgcacatttc cccgaaaagt gccagatacc tgaaacaaaa cccatcgtac ggccaaggaa
     3661 gtctccaata actgtgatcc accacaagcg ccagggtttt cccagtcacg acgttgtaaa
     3721 acgacggcca gtcatgcata atccgcacgc atctggaata aggaagtgcc attc
//
