LOCUS       sequence-367816-        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-730pUC19-tNGFR-P2A-Coronin1A
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1177..1233
                     /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(1611..1627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1611..1627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1624..1646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1799..2387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1879..1898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2504..2528)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2628..3419,3420..3488))
                     /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     3251..3270
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3489..3593)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3692..3714
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3706..3723
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3707..3723
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcgagctcg gtacccgggg atcaacaaac atgcagtggg gcaacaccac gccaggctct
       61 gttgcagagg ctgagggtac agattgagag ggtggctcag agtggcctgg ggccagggag
      121 aagggggctt gcaggttaga gggaaggaga ggctggggaa ggcaagcagg catcagccac
      181 agagggaaga atagggagcc cctggagatg atgctgggac cttaaaagcc agactgaggg
      241 gtgtcctggg ggaggtgggg accactggaa gatcagagtt gacctgaagg gagaaatgct
      301 cttatgctgt gtgcccccag aatgggtgct ggtgcaacgg ggagggctat ggatggacca
      361 aggctccttc ttcttttgct cttgggcgtt agtctgggtg gagctaaaga ggcatgcccc
      421 accggacttt acacacattc cggtgagtgt tgcaaggcct gcaacctggg agaaggcgtc
      481 gctcaaccct gcggggcgaa tcaaaccgta tgcgaacctt gcctcgattc tgttacattc
      541 tccgatgtcg tctctgcgac ggaaccctgc aagccatgca cggaatgtgt cgggttgcaa
      601 agtatgtcag ctccttgcgt tgaagcagac gatgcggtgt gtagatgtgc ctatgggtat
      661 tatcaagacg aaacgacggg acggtgtgaa gcatgtcgag tatgcgaggc gggcagtggc
      721 ctcgtcttct cttgtcagga caagcaaaac actgtctgtg aagaatgccc tgatgggact
      781 tatagtgatg aggcgaatca cgtcgatcca tgtcttccct gtactgtctg tgaagacact
      841 gagagacaat tgcgagagtg tacgcgatgg gccgacgcag agtgcgagga gatcccaggt
      901 cgatggataa ccaggtcaac ccctcctgag ggatctgata gtaccgcgcc ctccacccaa
      961 gaacccgaag ctccacctga gcaagacctc atagcgtcca cagtagcagg agttgtcaca
     1021 accgtcatgg gaagttcaca accagtagtt accaggggta ccacggataa cctcattccg
     1081 gtgtactgca gcattctcgc cgctgtggtt gtcggacttg ttgcatacat tgccttcaaa
     1141 cgctggaata gccgcgccaa gcgctcgggt tcgggtgcca ccaacttcag cctgctgaag
     1201 caggccggcg acgtggagga gaaccccggc cccagccggc aggtggtccg ctcgagcaag
     1261 ttccgccacg tgtttggaca gccggccaag gccgaccagt gctatgaaga tgtgcgcgtc
     1321 tcacagacca cctgggacag tggcttctgt gctgtcaacc ctaagtttgt ggccctgatc
     1381 tgtgaggcca gcgggggagg ggccttcctg gtgctgcccc tgggcaaggt gagcccctgg
     1441 ggccctgggg ggagcagctc ctccaccgga cccatggctc tgtgcaggtc ctgattaggt
     1501 gacggtcacc aggccttcca gggcctgtgt tgctcctcta gagtcgacct gcagaggcta
     1561 ggtggaggct cagtgatgat aagtctgcga tggtggatgc atgtgtcatg gtcatagctg
     1621 tttcctgtgt gaaattgtta tccgctcaga gggcacaatc ctattccgcg ctatccgaca
     1681 atctccaaga cattaggtgg agttcagttc ggcgtatggc atatgtcgct ggaaagaaca
     1741 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2221 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2341 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2401 tcttttctac ggggtctgac gctctattca acaaagccgc cgtcccgtca agtcagcgta
     2461 aatgggtagg gggcttcaaa tcgtcctcgt gataccaatt cggagcctgc ttttttgtac
     2521 aaacttgttg ataatggcaa ttcaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2581 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2641 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2701 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2761 ataccgcgag agccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2821 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2881 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2941 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3001 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3061 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3121 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3181 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3241 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3301 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3361 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3421 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3481 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3541 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3601 ccccgaaaag tgccagatac ctgaaacaaa acccatcgta cggccaagga agtctccaat
     3661 aactgtgatc caccacaagc gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     3721 agtcatgcat aatccgcacg catctggaat aaggaagtgc cattccgcct gacc
//
