LOCUS       sequence-367781-        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-728pUC19-tNGFR-P2A-WIP
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1161..1217
                     /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(1595..1611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1595..1611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1608..1630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1783..2371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1863..1882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2488..2512)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2612..3403,3404..3472))
                     /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     3235..3254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3473..3577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3676..3698
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3690..3707
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3691..3707
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggggaggcct actgtagtgg gtgggggaat ggagccgagt aaggcactgg atgtggcctc
       61 cattctctcc aagcacctcg ggtcagaagc ccctctgccg ggatccttcc agaacagtaa
      121 tgttttagca gtgaggtgta aaggtaagcc catctcttct aagagttgag ctaggtggag
      181 ctgtcacttt ggtgggaaag tgaagacaga ttaggacagt attactaaga ttacatctaa
      241 tacatgatcg cattcttgtt tttccagatt tatcagcaag actgttgaac gcataactgc
      301 ccaagatggg tgctggtgca acggggaggg ctatggatgg accaaggctc cttcttcttt
      361 tgctcttggg cgttagtctg ggtggagcta aagaggcatg ccccaccgga ctttacacac
      421 attccggtga gtgttgcaag gcctgcaacc tgggagaagg cgtcgctcaa ccctgcgggg
      481 cgaatcaaac cgtatgcgaa ccttgcctcg attctgttac attctccgat gtcgtctctg
      541 cgacggaacc ctgcaagcca tgcacggaat gtgtcgggtt gcaaagtatg tcagctcctt
      601 gcgttgaagc agacgatgcg gtgtgtagat gtgcctatgg gtattatcaa gacgaaacga
      661 cgggacggtg tgaagcatgt cgagtatgcg aggcgggcag tggcctcgtc ttctcttgtc
      721 aggacaagca aaacactgtc tgtgaagaat gccctgatgg gacttatagt gatgaggcga
      781 atcacgtcga tccatgtctt ccctgtactg tctgtgaaga cactgagaga caattgcgag
      841 agtgtacgcg atgggccgac gcagagtgcg aggagatccc aggtcgatgg ataaccaggt
      901 caacccctcc tgagggatct gatagtaccg cgccctccac ccaagaaccc gaagctccac
      961 ctgagcaaga cctcatagcg tccacagtag caggagttgt cacaaccgtc atgggaagtt
     1021 cacaaccagt agttaccagg ggtaccacgg ataacctcat tccggtgtac tgcagcattc
     1081 tcgccgctgt ggttgtcgga cttgttgcat acattgcctt caaacgctgg aatagccgcg
     1141 ccaagcgctc gggttcgggt gccaccaact tcagcctgct gaagcaggcc ggcgacgtgg
     1201 aggagaaccc cggcccccct gtgccccctc cacctgctcc cccgcccact cctacgtttg
     1261 cactggtgag tgtctcctcc ccaaacactt tctatgcata aagcaagcca aatgaagcca
     1321 aaacctggcg tgcatgtgtt tgtattaaat gtgtttgcat tctggctcac atgtgataca
     1381 acaggctttc ccagtgccca gggtaagcca aatcctgaaa cccagcaaat acaaacaaca
     1441 cgaagggttt tgtttgttta aggtctcgtg tgatggatag tacaagggtg gaatgtggct
     1501 gatgaggtgt tttctgctcc tctagagtcg acctgcagag gctaggtgga ggctcagtga
     1561 tgataagtct gcgatggtgg atgcatgtgt catggtcata gctgtttcct gtgtgaaatt
     1621 gttatccgct cagagggcac aatcctattc cgcgctatcc gacaatctcc aagacattag
     1681 gtggagttca gttcggcgta tggcatatgt cgctggaaag aacatgtgag caaaaggcca
     1741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2401 tgacgctcta ttcaacaaag ccgccgtccc gtcaagtcag cgtaaatggg tagggggctt
     2461 caaatcgtcc tcgtgatacc aattcggagc ctgctttttt gtacaaactt gttgataatg
     2521 gcaattcaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2581 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2641 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2701 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagagccac
     2761 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2821 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2881 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2941 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3001 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3061 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3121 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3181 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3241 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3301 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3361 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3421 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3481 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3541 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccag
     3601 atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt gatccaccac
     3661 aagcgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtcat gcataatccg
     3721 cacgcatctg gaataaggaa gtgccattcc gcctgacctt cgagctcggt accc
//
