LOCUS       sequence-367777-        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-716pUC19-tNGFR-P2A-CD3G
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1168..1224
                     /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(1602..1618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1602..1618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1615..1637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1790..2378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1870..1889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2495..2519)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2619..3410,3411..3479))
                     /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     3242..3261
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3480..3584)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3683..3705
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3697..3714
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3698..3714
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtacccggg gaccattgcg gttccctgtg caagatgagt ctctgagtgg gaatccagca
       61 ctctctccct cttcttcccc accaccttca ccctccttaa cggaaaaaca aaaggcatct
      121 gcacctgcag ccctgctgag gcccctgctg ctcacacttg cagcagaggg tggaggctct
      181 gggttcttgc cttctctcaa aggccccagc cccaacagtg atgggtggag ccagtctagc
      241 tgctgcacag gctggctggc tggctggctg ctaagggctg ctccacgctt ttgcgggagg
      301 acacagccta acatgggtgc tggtgcaacg gggagggcta tggatggacc aaggctcctt
      361 cttcttttgc tcttgggcgt tagtctgggt ggagctaaag aggcatgccc caccggactt
      421 tacacacatt ccggtgagtg ttgcaaggcc tgcaacctgg gagaaggcgt cgctcaaccc
      481 tgcggggcga atcaaaccgt atgcgaacct tgcctcgatt ctgttacatt ctccgatgtc
      541 gtctctgcga cggaaccctg caagccatgc acggaatgtg tcgggttgca aagtatgtca
      601 gctccttgcg ttgaagcaga cgatgcggtg tgtagatgtg cctatgggta ttatcaagac
      661 gaaacgacgg gacggtgtga agcatgtcga gtatgcgagg cgggcagtgg cctcgtcttc
      721 tcttgtcagg acaagcaaaa cactgtctgt gaagaatgcc ctgatgggac ttatagtgat
      781 gaggcgaatc acgtcgatcc atgtcttccc tgtactgtct gtgaagacac tgagagacaa
      841 ttgcgagagt gtacgcgatg ggccgacgca gagtgcgagg agatcccagg tcgatggata
      901 accaggtcaa cccctcctga gggatctgat agtaccgcgc cctccaccca agaacccgaa
      961 gctccacctg agcaagacct catagcgtcc acagtagcag gagttgtcac aaccgtcatg
     1021 ggaagttcac aaccagtagt taccaggggt accacggata acctcattcc ggtgtactgc
     1081 agcattctcg ccgctgtggt tgtcggactt gttgcataca ttgccttcaa acgctggaat
     1141 agccgcgcca agcgctcggg ttcgggtgcc accaacttca gcctgctgaa gcaggccggc
     1201 gacgtggagg agaaccccgg ccccgaacag gggaaaggcc tcgctgtcct catcctggct
     1261 atcattcttc ttcaaggtaa gggcctacta ggggtctgga agcctgggga agggctcaag
     1321 ggaagagccc atcactagtg agacaggaat attggtatcc ctaaccttca gcctacctct
     1381 gctgtcacct tagagttcaa agaagggcaa aatggaggct cttaactgtt ctctgctaga
     1441 gagaaacagt gtcccatgga ggagaaggaa tccttgtctc tgaaaaatgc aaacagagta
     1501 cttaaatggc tgaagagagg accctcctct agagtcgacc tgcagaggct aggtggaggc
     1561 tcagtgatga taagtctgcg atggtggatg catgtgtcat ggtcatagct gtttcctgtg
     1621 tgaaattgtt atccgctcag agggcacaat cctattccgc gctatccgac aatctccaag
     1681 acattaggtg gagttcagtt cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa
     1741 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1801 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1861 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1921 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1981 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2041 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2101 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2161 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2221 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2281 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2341 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2401 cggggtctga cgctctattc aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag
     2461 ggggcttcaa atcgtcctcg tgataccaat tcggagcctg cttttttgta caaacttgtt
     2521 gataatggca attcaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2581 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2641 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2701 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2761 gagccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2821 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2881 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2941 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3001 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3061 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3121 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3181 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3241 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3301 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3361 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3421 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3481 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3541 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3601 gtgccagata cctgaaacaa aacccatcgt acggccaagg aagtctccaa taactgtgat
     3661 ccaccacaag cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtcatgca
     3721 taatccgcac gcatctggaa taaggaagtg ccattccgcc tgaccttcga gctc
//
