LOCUS       sequence-367283-        4259 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 4259)
  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 4259)
  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: pTR-471 IRF4-tNGFR
FEATURES             Location/Qualifiers
     source          1..4259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..47)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     147..166
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     360..382
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     374..391
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     375..391
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1699..1755
                     /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(2034..2050)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2034..2050)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2047..2069)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2058..2074
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2082..2088,2089..2106,2107..2112))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2127..2148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2265..2282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2436..3024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2516..2535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3195..3986,3987..4055))
                     /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     3818..3837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4056..4160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4228..4246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
       61 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      121 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      181 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      241 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      301 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      361 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc ggggaaaggc
      421 aagcgagagc tgcgagtgag tgcggaagga gggccaggag gggtggcggc tgggtgggga
      481 gagagggtgc aagacgagcg gcgcgtgtcg ggagcctttg ggctgcgggt gcgttacagg
      541 agagcaggcg ggtaggagcc ttcgcggggg ccgagctcgg aaggcggacg gctgtgcccg
      601 cccaggggat gcgcccgggc cggccgcgaa ggtgccttct tccgggggcc cggacgaccc
      661 tgacacggca cgcgcgcgct tcgcagcctc aaagactccg gggcctcgtg gtcactggcg
      721 caggggatcg gggcggggtg cccggagtgc ggtgcctcgt ggctgaaggg cagctcttct
      781 ccccgcagtg cagagcagag cgggcggagg accccgggcg cgggcgcgga cggcacgcgg
      841 ggcatggggg caggtgccac tggccgcgcc atggacgggc cgcgcctgct gctgttgctg
      901 cttctggggg tgtcccttgg aggtgccaag gaggcatgcc ccacaggcct gtacacacac
      961 agcggtgagt gctgcaaagc ctgcaacctg ggcgagggtg tggcccagcc ttgtggagcc
     1021 aaccagaccg tgtgtgagcc ctgcctggac agcgtgacgt tctccgacgt ggtgagcgcg
     1081 accgagccgt gcaagccgtg caccgagtgc gtggggctcc agagcatgtc ggcgccatgc
     1141 gtggaggccg acgacgccgt gtgccgctgc gcctacggct actaccagga tgagacgact
     1201 gggcgctgcg aggcgtgccg cgtgtgcgag gcgggctcgg gcctcgtgtt ctcctgccag
     1261 gacaagcaga acaccgtgtg cgaggagtgc cccgacggca cgtattccga cgaggccaac
     1321 cacgtggacc cgtgcctgcc ctgcaccgtg tgcgaggaca ccgagcgcca gctccgcgag
     1381 tgcacacgct gggccgacgc cgagtgcgag gagatccctg gccgttggat tacacggtcc
     1441 acacccccag agggctcgga cagcacagcc cccagcaccc aggagcctga ggcacctcca
     1501 gaacaagacc tcatagccag cacggtggca ggtgtggtga ccacagtgat gggcagctcc
     1561 cagcccgtgg tgacccgagg caccaccgac aacctcatcc ctgtctattg ctccatcctg
     1621 gctgctgtgg ttgtgggtct tgtggcctac atagccttca agaggtggaa cagccgcgcc
     1681 aagcgctcgg gttcgggtgc caccaacttc agcctgctga agcaggccgg cgacgtggag
     1741 gagaaccctg gccctatgaa cctggaaggc ggcggccgag gcggagagtt cggcatgagc
     1801 gcggtgagct gcggcaacgg gaagctccgc cagtggctga tcgaccagat cgacagcggc
     1861 aagtaccccg ggctggtgtg ggagaacgag gagaagagca tcttccgcat cccctggaag
     1921 cacgcgggca agcaggacta caaccgcgag gaggacgccg cgctcttcaa ggtctccggc
     1981 ctcgggagtc ctctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag
     2041 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2101 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2161 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2221 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2281 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2341 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2401 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2461 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2521 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2581 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2641 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2701 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2761 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2821 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2881 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2941 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3001 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3061 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3121 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3181 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3241 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3301 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3361 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3421 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3481 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3541 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3601 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3661 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3721 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3781 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3841 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3901 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3961 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4021 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4081 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4141 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4201 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgc
//
