LOCUS       sequence-367780-        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-731pUC19-tNGFR-P2A-MAGT1
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1173..1229
                     /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"
     CDS             1302..1313
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1607..1623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1607..1623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1620..1642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1795..2383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1875..1894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2500..2524)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2624..3415,3416..3484))
                     /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     3247..3266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3485..3589)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3688..3710
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3702..3719
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3703..3719
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agctcggtac ccggggattc taaatgagtc tttgaaagat tctgttctag caacttcaag
       61 ccattttgga ctccgaaaac tccgcaatat ttcacaactg cgcatgcgtg cttggaaagc
      121 gtcagcgcgc tagatgttta cgtatcgaga aaggggcggg tcattatctt tagcggacca
      181 atgaaaacgc tccagattat cttttttcta ttggccgacc ggggccaatt agaaatcaac
      241 cttcttttag cgggcgtgta gcgccagcgc gctgtgacgt aatgtgaggg gtctcgccgc
      301 agggctgagc tcgaccaatg ggtgctggtg caacggggag ggctatggat ggaccaaggc
      361 tccttcttct tttgctcttg ggcgttagtc tgggtggagc taaagaggca tgccccaccg
      421 gactttacac acattccggt gagtgttgca aggcctgcaa cctgggagaa ggcgtcgctc
      481 aaccctgcgg ggcgaatcaa accgtatgcg aaccttgcct cgattctgtt acattctccg
      541 atgtcgtctc tgcgacggaa ccctgcaagc catgcacgga atgtgtcggg ttgcaaagta
      601 tgtcagctcc ttgcgttgaa gcagacgatg cggtgtgtag atgtgcctat gggtattatc
      661 aagacgaaac gacgggacgg tgtgaagcat gtcgagtatg cgaggcgggc agtggcctcg
      721 tcttctcttg tcaggacaag caaaacactg tctgtgaaga atgccctgat gggacttata
      781 gtgatgaggc gaatcacgtc gatccatgtc ttccctgtac tgtctgtgaa gacactgaga
      841 gacaattgcg agagtgtacg cgatgggccg acgcagagtg cgaggagatc ccaggtcgat
      901 ggataaccag gtcaacccct cctgagggat ctgatagtac cgcgccctcc acccaagaac
      961 ccgaagctcc acctgagcaa gacctcatag cgtccacagt agcaggagtt gtcacaaccg
     1021 tcatgggaag ttcacaacca gtagttacca ggggtaccac ggataacctc attccggtgt
     1081 actgcagcat tctcgccgct gtggttgtcg gacttgttgc atacattgcc ttcaaacgct
     1141 ggaatagccg cgccaagcgc tcgggttcgg gtgccaccaa cttcagcctg ctgaagcagg
     1201 ccggcgacgt ggaggagaac cccggcccca ggaaaggcaa ggggccgatt tgcctgttct
     1261 cacgccccac cctcagacct agccggagca aagtttcact tatagaaggg agaggagcga
     1321 acatggcagc gcgttggcgg ttttggtgtg tctctgtgac catggtggtg gcgctgctca
     1381 tcgtttgcga cgttccctca gcctctgccc aaagaaagaa ggaggtgaga acgcggtttc
     1441 cagcagcatg ggcttttccc aatgactggg gcttaagagg gtctgttcgc ctcttcccag
     1501 ccccctttcc ctgccgcttc tatgcctttt cctctagagt cgacctgcag aggctaggtg
     1561 gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca tagctgtttc
     1621 ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat ccgacaatct
     1681 ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa agaacatgtg
     1741 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1801 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1861 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1921 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1981 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2041 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2101 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2161 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2221 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2281 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2341 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2401 ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc agcgtaaatg
     2461 ggtagggggc ttcaaatcgt cctcgtgata ccaattcgga gcctgctttt ttgtacaaac
     2521 ttgttgataa tggcaattca aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2581 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2641 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2701 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2761 cgcgagagcc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2821 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2881 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     2941 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3001 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3061 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3121 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3181 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3241 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3301 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3361 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3421 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3481 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3541 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3601 gaaaagtgcc agatacctga aacaaaaccc atcgtacggc caaggaagtc tccaataact
     3661 gtgatccacc acaagcgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtc
     3721 atgcataatc cgcacgcatc tggaataagg aagtgccatt ccgcctgacc ttcg
//
