LOCUS       sequence-367804-        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-705pUC19-tNGFR-P2A-STAT5b
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1189..1245
                     /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(1623..1639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1623..1639)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1636..1658)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1811..2399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1891..1910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2516..2540)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2640..3431,3432..3500))
                     /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     3263..3282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3501..3605)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3704..3726
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3718..3735
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3719..3735
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttccgcctga ccttcgagct cggtacccgg ggaaaaatac aaaaattaac caagcatggt
       61 agtgtgcccc tgtaatccca gctactcagg aggctgagac aggagaatca tttgaaccgg
      121 ggaggcggag gttgtagtga gccaagattg tgccactgca ctccagcttg ggtggcagag
      181 cgagactgtc tcaacgaagt aagaataaga atacataata gaataacaac atttaccttg
      241 gtaactaagt gaatatttac tctggtggat gtaaggggaa aataaagaaa aaacgcccaa
      301 agcactgatt gtttgttcat cagaatggaa gccatgggtg ctggtgcaac ggggagggct
      361 atggatggac caaggctcct tcttcttttg ctcttgggcg ttagtctggg tggagctaaa
      421 gaggcatgcc ccaccggact ttacacacat tccggtgagt gttgcaaggc ctgcaacctg
      481 ggagaaggcg tcgctcaacc ctgcggggcg aatcaaaccg tatgcgaacc ttgcctcgat
      541 tctgttacat tctccgatgt cgtctctgcg acggaaccct gcaagccatg cacggaatgt
      601 gtcgggttgc aaagtatgtc agctccttgc gttgaagcag acgatgcggt gtgtagatgt
      661 gcctatgggt attatcaaga cgaaacgacg ggacggtgtg aagcatgtcg agtatgcgag
      721 gcgggcagtg gcctcgtctt ctcttgtcag gacaagcaaa acactgtctg tgaagaatgc
      781 cctgatggga cttatagtga tgaggcgaat cacgtcgatc catgtcttcc ctgtactgtc
      841 tgtgaagaca ctgagagaca attgcgagag tgtacgcgat gggccgacgc agagtgcgag
      901 gagatcccag gtcgatggat aaccaggtca acccctcctg agggatctga tagtaccgcg
      961 ccctccaccc aagaacccga agctccacct gagcaagacc tcatagcgtc cacagtagca
     1021 ggagttgtca caaccgtcat gggaagttca caaccagtag ttaccagggg taccacggat
     1081 aacctcattc cggtgtactg cagcattctc gccgctgtgg ttgtcggact tgttgcatac
     1141 attgccttca aacgctggaa tagccgcgcc aagcgctcgg gttcgggtgc caccaacttc
     1201 agcctgctga agcaggccgg cgacgtggag gagaaccccg gccccgctgt gtggatacaa
     1261 gctcaacaac tgcagggaga agcccttcat cagatgcaag cgttatatgg ccagcatttt
     1321 cccattgagg tgcggcatta tttatcccag tggattgaaa gccaagcatg gtaggtgaac
     1381 aaggatgttt gcatcatatt ttgtgaaaca caagaatatt ggagtttgtt tagaaagatg
     1441 tcatgatggc aactttcaaa gttgtatttt gaaacattgt taaatgtgat ccataaacag
     1501 gtattgaaaa tgtttcttct ttttataggt gaaaggtatt taagctcctc tagagtcgac
     1561 ctgcagaggc taggtggagg ctcagtgatg ataagtctgc gatggtggat gcatgtgtca
     1621 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa tcctattccg
     1681 cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg gcatatgtcg
     1741 ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2221 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2341 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2401 aagatccttt gatcttttct acggggtctg acgctctatt caacaaagcc gccgtcccgt
     2461 caagtcagcg taaatgggta gggggcttca aatcgtcctc gtgataccaa ttcggagcct
     2521 gcttttttgt acaaacttgt tgataatggc aattcaagga tcttcaccta gatcctttta
     2581 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2641 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2701 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2761 agtgctgcaa tgataccgcg agagccacgc tcaccggctc cagatttatc agcaataaac
     2821 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2881 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2941 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3001 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3061 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3121 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3181 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3241 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3301 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3361 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3421 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3481 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3541 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3601 ccgcgcacat ttccccgaaa agtgccagat acctgaaaca aaacccatcg tacggccaag
     3661 gaagtctcca ataactgtga tccaccacaa gcgccagggt tttcccagtc acgacgttgt
     3721 aaaacgacgg ccagtcatgc ataatccgca cgcatctgga ataaggaagt gcca
//
