LOCUS       sequence-367514-        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-747pUC19-tNGFR-P2A-CD7
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      323..332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1184..1240
                     /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(1618..1634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1618..1634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1631..1653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1806..2394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1886..1905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2511..2535)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2635..3426,3427..3495))
                     /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     3258..3277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3496..3600)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3699..3721
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3713..3730
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3714..3730
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cctgaccttc gagctcggta cccggggact ctggtctgcc ccgccccagg ccttgacctc
       61 ctccctgtgg agatgcaggg gcagtggggg atacagggac gccctgctct cagggcagct
      121 ctcagggagg cagtgctggg gaggggtagg tgagaccgcc cctcccaccg ggcccacagc
      181 accaccctgg tctgaggcac cgcctccagg aagccctctc tgagctctga gcgcctgcgg
      241 tctcctgtgt gctgctctct gtggggtcct gtagacccag agaggctcag ctgcactcgc
      301 ccggctgcga gagctgggag tggccaccat gggtgctggt gcaacgggga gggctatgga
      361 tggaccaagg ctccttcttc ttttgctctt gggcgttagt ctgggtggag ctaaagaggc
      421 atgccccacc ggactttaca cacattccgg tgagtgttgc aaggcctgca acctgggaga
      481 aggcgtcgct caaccctgcg gggcgaatca aaccgtatgc gaaccttgcc tcgattctgt
      541 tacattctcc gatgtcgtct ctgcgacgga accctgcaag ccatgcacgg aatgtgtcgg
      601 gttgcaaagt atgtcagctc cttgcgttga agcagacgat gcggtgtgta gatgtgccta
      661 tgggtattat caagacgaaa cgacgggacg gtgtgaagca tgtcgagtat gcgaggcggg
      721 cagtggcctc gtcttctctt gtcaggacaa gcaaaacact gtctgtgaag aatgccctga
      781 tgggacttat agtgatgagg cgaatcacgt cgatccatgt cttccctgta ctgtctgtga
      841 agacactgag agacaattgc gagagtgtac gcgatgggcc gacgcagagt gcgaggagat
      901 cccaggtcga tggataacca ggtcaacccc tcctgaggga tctgatagta ccgcgccctc
      961 cacccaagaa cccgaagctc cacctgagca agacctcata gcgtccacag tagcaggagt
     1021 tgtcacaacc gtcatgggaa gttcacaacc agtagttacc aggggtacca cggataacct
     1081 cattccggtg tactgcagca ttctcgccgc tgtggttgtc ggacttgttg catacattgc
     1141 cttcaaacgc tggaatagcc gcgccaagcg ctcgggttcg ggtgccacca acttcagcct
     1201 gctgaagcag gccggcgacg tggaggagaa ccccggcccc gccgggcctc cgagactgct
     1261 cctcctgccc ctgcttctgg cgctggctcg cggcctgcct ggggccctgg ctgcccaagg
     1321 taagagcttc ccaggctctc catggccaca gctccagagc tctccctgcc ccatgagctc
     1381 agagccccca gtctgagcca cagcacagcc cccaggaagc gggtggggtg ctgagcggcc
     1441 tccagtgtct gaggactcat ttaagagaag gaaaaagggt ggaccccggt ggggagtggc
     1501 cggggctgtc caggcagggc cgctgctttg ggaggaagaa tcctctagag tcgacctgca
     1561 gaggctaggt ggaggctcag tgatgataag tctgcgatgg tggatgcatg tgtcatggtc
     1621 atagctgttt cctgtgtgaa attgttatcc gctcagaggg cacaatccta ttccgcgcta
     1681 tccgacaatc tccaagacat taggtggagt tcagttcggc gtatggcata tgtcgctgga
     1741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2401 cctttgatct tttctacggg gtctgacgct ctattcaaca aagccgccgt cccgtcaagt
     2461 cagcgtaaat gggtaggggg cttcaaatcg tcctcgtgat accaattcgg agcctgcttt
     2521 tttgtacaaa cttgttgata atggcaattc aaggatcttc acctagatcc ttttaaatta
     2581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2701 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2761 tgcaatgata ccgcgagagc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2821 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2881 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2941 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3001 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3061 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3421 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3481 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3601 cacatttccc cgaaaagtgc cagatacctg aaacaaaacc catcgtacgg ccaaggaagt
     3661 ctccaataac tgtgatccac cacaagcgcc agggttttcc cagtcacgac gttgtaaaac
     3721 gacggccagt catgcataat ccgcacgcat ctggaataag gaagtgccat tccg
//
