LOCUS       sequence-367817-        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-717pUC19-tNGFR-P2A-CD3E
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1172..1228
                     /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(1606..1622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1606..1622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1619..1641)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1794..2382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1874..1893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2499..2523)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2623..3414,3415..3483))
                     /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     3246..3265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3484..3588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3687..3709
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3701..3718
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3702..3718
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gctcggtacc cggggaaggc ctctgccttg aacgtttcca agtgaggtaa aacccgcagg
       61 cccagaggcc tctctacttc ctgtgtgggg ttcagaaacc ctcctcccct cccagcctca
      121 ggtgcctgct tcagaaaatg gtgagtctct ctcttataaa gccctccttt ttcatcctag
      181 cattgggaac aatggcccca gggtccttat ctctagcaga tgttttgaaa aagtcatctg
      241 ttttgctttt tttccagaag tagtaagtct gctggcctcc gccatcttag taaagtaaca
      301 gtcgcatgaa acaaagatgg gtgctggtgc aacggggagg gctatggatg gaccaaggct
      361 ccttcttctt ttgctcttgg gcgttagtct gggtggagct aaagaggcat gccccaccgg
      421 actttacaca cattccggtg agtgttgcaa ggcctgcaac ctgggagaag gcgtcgctca
      481 accctgcggg gcgaatcaaa ccgtatgcga accttgcctc gattctgtta cattctccga
      541 tgtcgtctct gcgacggaac cctgcaagcc atgcacggaa tgtgtcgggt tgcaaagtat
      601 gtcagctcct tgcgttgaag cagacgatgc ggtgtgtaga tgtgcctatg ggtattatca
      661 agacgaaacg acgggacggt gtgaagcatg tcgagtatgc gaggcgggca gtggcctcgt
      721 cttctcttgt caggacaagc aaaacactgt ctgtgaagaa tgccctgatg ggacttatag
      781 tgatgaggcg aatcacgtcg atccatgtct tccctgtact gtctgtgaag acactgagag
      841 acaattgcga gagtgtacgc gatgggccga cgcagagtgc gaggagatcc caggtcgatg
      901 gataaccagg tcaacccctc ctgagggatc tgatagtacc gcgccctcca cccaagaacc
      961 cgaagctcca cctgagcaag acctcatagc gtccacagta gcaggagttg tcacaaccgt
     1021 catgggaagt tcacaaccag tagttaccag gggtaccacg gataacctca ttccggtgta
     1081 ctgcagcatt ctcgccgctg tggttgtcgg acttgttgca tacattgcct tcaaacgctg
     1141 gaatagccgc gccaagcgct cgggttcggg tgccaccaac ttcagcctgc tgaagcaggc
     1201 cggcgacgtg gaggagaacc ccggccccca gtccgggaca cattggaggg tattgggcct
     1261 ctgcctctta tcaggtgagt aggatggagt ggaaagggtg gtgtgtctcc agaccgctgg
     1321 aaggcttaca gccttacctg gcactgccta gtggcaccaa ggagcctcat ttaccagatg
     1381 taaggaactg tttgtgctat gttagggtga gggattagag ctggggacta aagaaaaaga
     1441 taggccacgg gtgcctggga gagcgttcgg ggagcaggca aagaagagca gttggggtga
     1501 tcatagctat tgtgagcaga gaggtctctc ctctagagtc gacctgcaga ggctaggtgg
     1561 aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc
     1621 tgtgtgaaat tgttatccgc tcagagggca caatcctatt ccgcgctatc cgacaatctc
     1681 caagacatta ggtggagttc agttcggcgt atggcatatg tcgctggaaa gaacatgtga
     1741 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1801 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1861 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1921 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1981 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2041 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2101 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2161 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2221 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2281 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2401 tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca gcgtaaatgg
     2461 gtagggggct tcaaatcgtc ctcgtgatac caattcggag cctgcttttt tgtacaaact
     2521 tgttgataat ggcaattcaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2761 gcgagagcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3601 aaaagtgcca gatacctgaa acaaaaccca tcgtacggcc aaggaagtct ccaataactg
     3661 tgatccacca caagcgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtca
     3721 tgcataatcc gcacgcatct ggaataagga agtgccattc cgcctgacct tcga
//
