LOCUS       sequence-367796-        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-742pUC19-tNGFR-P2A-Caspase8
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1175..1231
                     /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(1609..1625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1609..1625)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1622..1644)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1797..2385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1877..1896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2502..2526)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2626..3417,3418..3486))
                     /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     3249..3268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3487..3591)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3690..3712
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3704..3721
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3705..3721
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgagctcggt acccggggag gctttagttt gcacgtccat gaattgtctg ccacatccct
       61 cttctgaatg gttggaaatt gggcatctgt tcctttaaac aggaaacatt tcttgttcga
      121 gtgagtcatc tctgttctgc tttaggagta aagtttaccc tgcagttcct tctgtggtga
      181 agttttctct ttctctcgga gaccagattc tgcctttctg ctggagggaa gtgttttcac
      241 aggttctcct ccttttatct tttgtgtttt ttttcaagcc ctgctgaatt tgctagtcaa
      301 ctcaacagga agtgacgcca tgggtgctgg tgcaacgggg agggctatgg atggaccaag
      361 gctccttctt cttttgctct tgggcgttag tctgggtgga gctaaagagg catgccccac
      421 cggactttac acacattccg gtgagtgttg caaggcctgc aacctgggag aaggcgtcgc
      481 tcaaccctgc ggggcgaatc aaaccgtatg cgaaccttgc ctcgattctg ttacattctc
      541 cgatgtcgtc tctgcgacgg aaccctgcaa gccatgcacg gaatgtgtcg ggttgcaaag
      601 tatgtcagct ccttgcgttg aagcagacga tgcggtgtgt agatgtgcct atgggtatta
      661 tcaagacgaa acgacgggac ggtgtgaagc atgtcgagta tgcgaggcgg gcagtggcct
      721 cgtcttctct tgtcaggaca agcaaaacac tgtctgtgaa gaatgccctg atgggactta
      781 tagtgatgag gcgaatcacg tcgatccatg tcttccctgt actgtctgtg aagacactga
      841 gagacaattg cgagagtgta cgcgatgggc cgacgcagag tgcgaggaga tcccaggtcg
      901 atggataacc aggtcaaccc ctcctgaggg atctgatagt accgcgccct ccacccaaga
      961 acccgaagct ccacctgagc aagacctcat agcgtccaca gtagcaggag ttgtcacaac
     1021 cgtcatggga agttcacaac cagtagttac caggggtacc acggataacc tcattccggt
     1081 gtactgcagc attctcgccg ctgtggttgt cggacttgtt gcatacattg ccttcaaacg
     1141 ctggaatagc cgcgccaagc gctcgggttc gggtgccacc aacttcagcc tgctgaagca
     1201 ggccggcgac gtggaggaga accccggccc cgagggaggc agaagagcga gagtggttat
     1261 tgaaagtaaa agaaacttct tcctgggagc ctttcccacc cccttccctg ctgagcacgt
     1321 ggagttaggc aggttagggg actcggagac tgcgatggtg ccaggaaagg gtggagcggg
     1381 tgagtgcctg ttgccaaggt ggcctcttca acaggaaacc acaatatttt tgtttcttga
     1441 cttgctctag aaacagggct gtgggggtgg ggaagcaact tggatctgcc cttctgagga
     1501 cacctctggg tgctgcctgg cccaggtctc ctcctctaga gtcgacctgc agaggctagg
     1561 tggaggctca gtgatgataa gtctgcgatg gtggatgcat gtgtcatggt catagctgtt
     1621 tcctgtgtga aattgttatc cgctcagagg gcacaatcct attccgcgct atccgacaat
     1681 ctccaagaca ttaggtggag ttcagttcgg cgtatggcat atgtcgctgg aaagaacatg
     1741 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1801 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1861 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1921 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1981 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2041 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2101 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2161 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2221 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2281 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2341 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2401 ttttctacgg ggtctgacgc tctattcaac aaagccgccg tcccgtcaag tcagcgtaaa
     2461 tgggtagggg gcttcaaatc gtcctcgtga taccaattcg gagcctgctt ttttgtacaa
     2521 acttgttgat aatggcaatt caaggatctt cacctagatc cttttaaatt aaaaatgaag
     2581 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2641 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2701 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2761 accgcgagag ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2821 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2881 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2941 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3001 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3061 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3121 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3181 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3241 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3301 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3361 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3421 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3481 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3541 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3601 ccgaaaagtg ccagatacct gaaacaaaac ccatcgtacg gccaaggaag tctccaataa
     3661 ctgtgatcca ccacaagcgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     3721 tcatgcataa tccgcacgca tctggaataa ggaagtgcca ttccgcctga cctt
//
