LOCUS       sequence-367776-        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-727pUC19-tNGFR-P2A-WASP
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1182..1238
                     /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(1616..1632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1616..1632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1629..1651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1804..2392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1884..1903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2509..2533)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2633..3424,3425..3493))
                     /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     3256..3275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3494..3598)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3697..3719
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3711..3728
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3712..3728
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgaccttcga gctcggtacc cggggagagg gtctgagcca gtcagaagga gatgggcccc
       61 agagagtaag aaagggggag gaggacccaa gctgatccaa aaggtgggtc taagcagtca
      121 agtggaggag ggttccaatc tgatggcgga gggcccaagc tcagcctaac gaggaggcca
      181 ggcccaccaa ggggcccctg gaggacttgt ttcccttgtc ccttgtggtt ttttgcattt
      241 cctgttccct tgctgctcat tgcggaagtt cctcttctta ccctgcaccc agagcctcgc
      301 gagagaagac aagggcagaa agcaccatgg gtgctggtgc aacggggagg gctatggatg
      361 gaccaaggct ccttcttctt ttgctcttgg gcgttagtct gggtggagct aaagaggcat
      421 gccccaccgg actttacaca cattccggtg agtgttgcaa ggcctgcaac ctgggagaag
      481 gcgtcgctca accctgcggg gcgaatcaaa ccgtatgcga accttgcctc gattctgtta
      541 cattctccga tgtcgtctct gcgacggaac cctgcaagcc atgcacggaa tgtgtcgggt
      601 tgcaaagtat gtcagctcct tgcgttgaag cagacgatgc ggtgtgtaga tgtgcctatg
      661 ggtattatca agacgaaacg acgggacggt gtgaagcatg tcgagtatgc gaggcgggca
      721 gtggcctcgt cttctcttgt caggacaagc aaaacactgt ctgtgaagaa tgccctgatg
      781 ggacttatag tgatgaggcg aatcacgtcg atccatgtct tccctgtact gtctgtgaag
      841 acactgagag acaattgcga gagtgtacgc gatgggccga cgcagagtgc gaggagatcc
      901 caggtcgatg gataaccagg tcaacccctc ctgagggatc tgatagtacc gcgccctcca
      961 cccaagaacc cgaagctcca cctgagcaag acctcatagc gtccacagta gcaggagttg
     1021 tcacaaccgt catgggaagt tcacaaccag tagttaccag gggtaccacg gataacctca
     1081 ttccggtgta ctgcagcatt ctcgccgctg tggttgtcgg acttgttgca tacattgcct
     1141 tcaaacgctg gaatagccgc gccaagcgct cgggttcggg tgccaccaac ttcagcctgc
     1201 tgaagcaggc cggcgacgtg gaggagaacc ccggccccag tggtggacct atggggggga
     1261 gacctggggg ccgaggagca ccagcggttc agcagaacat accctccacc ctcctccagg
     1321 accacgagaa ccagcgactc tttgagatgc ttggacgaaa atgcttggtg agctggggat
     1381 ctcctgcccc cgccccgtcc ccaccgtttc ttcctcttcc tctcctcctt ctctctcttc
     1441 ccctcctccc gctcctcctt tccctctcca tcatctcctc tcctagaatt tcccgtcata
     1501 atccaccctt cccaggaaga tctcaatgtc tacttgcctc ctctagagtc gacctgcaga
     1561 ggctaggtgg aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat
     1621 agctgtttcc tgtgtgaaat tgttatccgc tcagagggca caatcctatt ccgcgctatc
     1681 cgacaatctc caagacatta ggtggagttc agttcggcgt atggcatatg tcgctggaaa
     1741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2401 tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca
     2461 gcgtaaatgg gtagggggct tcaaatcgtc ctcgtgatac caattcggag cctgcttttt
     2521 tgtacaaact tgttgataat ggcaattcaa ggatcttcac ctagatcctt ttaaattaaa
     2581 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2641 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2701 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2761 caatgatacc gcgagagcca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2821 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2881 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2941 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3001 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3061 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3121 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3181 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3241 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3301 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3361 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3421 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3481 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3601 catttccccg aaaagtgcca gatacctgaa acaaaaccca tcgtacggcc aaggaagtct
     3661 ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt tgtaaaacga
     3721 cggccagtca tgcataatcc gcacgcatct ggaataagga agtgccattc cgcc
//
