LOCUS       sequence-367799-        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-740pUC19-tNGFR-P2A-BCL10
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1181..1237
                     /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(1615..1631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1615..1631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1628..1650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1803..2391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1883..1902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2508..2532)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2632..3423,3424..3492))
                     /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     3255..3274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3493..3597)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3696..3718
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3710..3727
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3711..3727
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaccttcgag ctcggtaccc ggggacgcgc ctcgccgtgg ggcggtgcct gcgcctgagc
       61 ctctacgaga gggaaggaac gctgctccga gctccgcgtc gcgtcgcgta gattcgcgtc
      121 gccgtcgacc tcagaggcgg ggccggaagc gctacggttt gacccccgag tccctctgtt
      181 cccgaagggg cggccgtctt tctcccgacc cgctccgcct cctctccttc ttccccatta
      241 cccggaggcc gaagccccca gccagggcgg ggcggcgcag cccgagctcc cggacccgga
      301 agaagcgcca tctcccgcct ccaccatggg tgctggtgca acggggaggg ctatggatgg
      361 accaaggctc cttcttcttt tgctcttggg cgttagtctg ggtggagcta aagaggcatg
      421 ccccaccgga ctttacacac attccggtga gtgttgcaag gcctgcaacc tgggagaagg
      481 cgtcgctcaa ccctgcgggg cgaatcaaac cgtatgcgaa ccttgcctcg attctgttac
      541 attctccgat gtcgtctctg cgacggaacc ctgcaagcca tgcacggaat gtgtcgggtt
      601 gcaaagtatg tcagctcctt gcgttgaagc agacgatgcg gtgtgtagat gtgcctatgg
      661 gtattatcaa gacgaaacga cgggacggtg tgaagcatgt cgagtatgcg aggcgggcag
      721 tggcctcgtc ttctcttgtc aggacaagca aaacactgtc tgtgaagaat gccctgatgg
      781 gacttatagt gatgaggcga atcacgtcga tccatgtctt ccctgtactg tctgtgaaga
      841 cactgagaga caattgcgag agtgtacgcg atgggccgac gcagagtgcg aggagatccc
      901 aggtcgatgg ataaccaggt caacccctcc tgagggatct gatagtaccg cgccctccac
      961 ccaagaaccc gaagctccac ctgagcaaga cctcatagcg tccacagtag caggagttgt
     1021 cacaaccgtc atgggaagtt cacaaccagt agttaccagg ggtaccacgg ataacctcat
     1081 tccggtgtac tgcagcattc tcgccgctgt ggttgtcgga cttgttgcat acattgcctt
     1141 caaacgctgg aatagccgcg ccaagcgctc gggttcgggt gccaccaact tcagcctgct
     1201 gaagcaggcc ggcgacgtgg aggagaaccc cggccccgag ccgaccgcac cgagtctgac
     1261 ggaggaggac ctcactgaag tgaagaagga cgtgagtaac gcagctgtgc ccagggcggg
     1321 cgggggcggg ctgcagccca gcgggagacg aaagcggaag cctggagtcc gaggacaagg
     1381 aggatcctcc aggtcggagg agcggaaagt cctagcacag gaggactgtg gcgagccctg
     1441 catccgaggg accttggtgg cagtgatcct ccagtgatct gtcaatccag gttttacatc
     1501 gctaaacgca gagcttgggc tttgttgcca agtggtgtcc tctagagtcg acctgcagag
     1561 gctaggtgga ggctcagtga tgataagtct gcgatggtgg atgcatgtgt catggtcata
     1621 gctgtttcct gtgtgaaatt gttatccgct cagagggcac aatcctattc cgcgctatcc
     1681 gacaatctcc aagacattag gtggagttca gttcggcgta tggcatatgt cgctggaaag
     1741 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1801 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1861 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1921 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1981 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2041 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2101 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2161 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2221 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2281 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2341 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2401 ttgatctttt ctacggggtc tgacgctcta ttcaacaaag ccgccgtccc gtcaagtcag
     2461 cgtaaatggg tagggggctt caaatcgtcc tcgtgatacc aattcggagc ctgctttttt
     2521 gtacaaactt gttgataatg gcaattcaag gatcttcacc tagatccttt taaattaaaa
     2581 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2641 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2701 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2761 aatgataccg cgagagccac gctcaccggc tccagattta tcagcaataa accagccagc
     2821 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2881 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2941 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3001 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3061 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3121 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3181 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3241 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3301 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3361 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3421 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3481 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3541 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3601 atttccccga aaagtgccag atacctgaaa caaaacccat cgtacggcca aggaagtctc
     3661 caataactgt gatccaccac aagcgccagg gttttcccag tcacgacgtt gtaaaacgac
     3721 ggccagtcat gcataatccg cacgcatctg gaataaggaa gtgccattcc gcct
//
