LOCUS       sequence-367828-        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-732pUC19-tNGFR-P2A-STIM1
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1170..1226
                     /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(1604..1620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1604..1620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1617..1639)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1792..2380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1872..1891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2497..2521)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2621..3412,3413..3481))
                     /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     3244..3263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3482..3586)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3685..3707
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3699..3716
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3700..3716
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcggtacccg gggagagctg acagcagccc cgcagccacc ctgcccgaag tctccggaag
       61 cggcacgagc tcaggccgcc gcagccccgg cggacccact gttggacctg aggagccagc
      121 cctcctcccg cacccaaact tggagcactt gacctttggc tgttggaggg ggcaggctcg
      181 cgggtggctg gacagctgcg gagccgcgag ggcatcttgc ctggagaccg tcggctgcac
      241 tcccgggctc ctggctttgc ctctgggatc ccgaggtgtc gacatcagac gcatgttgac
      301 tgagacctag agtcatgggt gctggtgcaa cggggagggc tatggatgga ccaaggctcc
      361 ttcttctttt gctcttgggc gttagtctgg gtggagctaa agaggcatgc cccaccggac
      421 tttacacaca ttccggtgag tgttgcaagg cctgcaacct gggagaaggc gtcgctcaac
      481 cctgcggggc gaatcaaacc gtatgcgaac cttgcctcga ttctgttaca ttctccgatg
      541 tcgtctctgc gacggaaccc tgcaagccat gcacggaatg tgtcgggttg caaagtatgt
      601 cagctccttg cgttgaagca gacgatgcgg tgtgtagatg tgcctatggg tattatcaag
      661 acgaaacgac gggacggtgt gaagcatgtc gagtatgcga ggcgggcagt ggcctcgtct
      721 tctcttgtca ggacaagcaa aacactgtct gtgaagaatg ccctgatggg acttatagtg
      781 atgaggcgaa tcacgtcgat ccatgtcttc cctgtactgt ctgtgaagac actgagagac
      841 aattgcgaga gtgtacgcga tgggccgacg cagagtgcga ggagatccca ggtcgatgga
      901 taaccaggtc aacccctcct gagggatctg atagtaccgc gccctccacc caagaacccg
      961 aagctccacc tgagcaagac ctcatagcgt ccacagtagc aggagttgtc acaaccgtca
     1021 tgggaagttc acaaccagta gttaccaggg gtaccacgga taacctcatt ccggtgtact
     1081 gcagcattct cgccgctgtg gttgtcggac ttgttgcata cattgccttc aaacgctgga
     1141 atagccgcgc caagcgctcg ggttcgggtg ccaccaactt cagcctgctg aagcaggccg
     1201 gcgacgtgga ggagaacccc ggccccgatg tatgcgtgcg tcttgctctg tggctcctct
     1261 ggggactcct cctgcaccag ggccagagcc tcagccatag tcacagtgag aaggcgacag
     1321 gaaccagctc gggggccaac tctgaggagt ccactgcagc aggtaaggcc ttgctgcggg
     1381 ctggactggg ctggaggctt tggctcagga ctgagtggcc cgaagtgggc aagttgaaat
     1441 ctaggtttgt acatgtgagg ttcatggagg attcacacat ggcactgcct gttccaagta
     1501 gttcaagaag ttctttctgg tcagtatcct ctagagtcga cctgcagagg ctaggtggag
     1561 gctcagtgat gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg
     1621 tgtgaaattg ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca
     1681 agacattagg tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc
     1741 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1801 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1861 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1921 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1981 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2041 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2101 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2161 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2221 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2281 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2341 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2401 tacggggtct gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt
     2461 agggggcttc aaatcgtcct cgtgatacca attcggagcc tgcttttttg tacaaacttg
     2521 ttgataatgg caattcaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2581 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2641 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2701 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2761 gagagccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2821 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2881 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2941 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3001 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3061 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3121 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3181 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3241 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3301 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3361 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3421 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3481 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3541 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3601 aagtgccaga tacctgaaac aaaacccatc gtacggccaa ggaagtctcc aataactgtg
     3661 atccaccaca agcgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtcatg
     3721 cataatccgc acgcatctgg aataaggaag tgccattccg cctgaccttc gagc
//
