LOCUS       sequence-367286-        4240 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 4240)
  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 4240)
  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-473 FOXP1-tNGFR
FEATURES             Location/Qualifiers
     source          1..4240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..60)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     160..179
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     373..395
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     387..404
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     388..404
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1503..1559
                     /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(2028..2044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2028..2044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2041..2063)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2052..2068
                     /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(2076..2082,2083..2100,2101..2106))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2121..2142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2259..2276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2430..3018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2510..2529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3189..3980,3981..4049))
                     /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     3812..3831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4050..4154)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4222..4240
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
       61 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      121 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      181 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      241 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      301 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      361 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta
      421 cccggggatt gaattgggaa tgacagtttc agacccgaaa ctaggggcat ggcccactaa
      481 tgaggggata agttgagtga aagaaaatga caactgttta cagattttgg caacatttaa
      541 accaaggccc ttctccttat gcacaacaac tgctttaaca gctgcttttt ttttctcccc
      601 cccaccctcc ccccatcttg gaaatccttg tatcaggttt ttgagtcatg ggggcaggtg
      661 ccactggccg cgccatggac gggccgcgcc tgctgctgtt gctgcttctg ggggtgtccc
      721 ttggaggtgc caaggaggca tgccccacag gcctgtacac acacagcggt gagtgctgca
      781 aagcctgcaa cctgggcgag ggtgtggccc agccttgtgg agccaaccag accgtgtgtg
      841 agccctgcct ggacagcgtg acgttctccg acgtggtgag cgcgaccgag ccgtgcaagc
      901 cgtgcaccga gtgcgtgggg ctccagagca tgtcggcgcc atgcgtggag gccgacgacg
      961 ccgtgtgccg ctgcgcctac ggctactacc aggatgagac gactgggcgc tgcgaggcgt
     1021 gccgcgtgtg cgaggcgggc tcgggcctcg tgttctcctg ccaggacaag cagaacaccg
     1081 tgtgcgagga gtgccccgac ggcacgtatt ccgacgaggc caaccacgtg gacccgtgcc
     1141 tgccctgcac cgtgtgcgag gacaccgagc gccagctccg cgagtgcaca cgctgggccg
     1201 acgccgagtg cgaggagatc cctggccgtt ggattacacg gtccacaccc ccagagggct
     1261 cggacagcac agcccccagc acccaggagc ctgaggcacc tccagaacaa gacctcatag
     1321 ccagcacggt ggcaggtgtg gtgaccacag tgatgggcag ctcccagccc gtggtgaccc
     1381 gaggcaccac cgacaacctc atccctgtct attgctccat cctggctgct gtggttgtgg
     1441 gtcttgtggc ctacatagcc ttcaagaggt ggaacagccg cgccaagcgc tcgggttcgg
     1501 gtgccaccaa cttcagcctg ctgaagcagg ccggcgacgt ggaggagaac cctggcccta
     1561 tgatgcaaga atctggcact gagacaaaaa gtaacggttc agccatccag aatgggtcgg
     1621 gcggcagcaa ccacttacta gagtgcggcg gtcttcggga ggggcggtcc aacggagaga
     1681 cgccggccgt ggacatcggg gcagctgacc tcgcccacgc ccagcagcag cagcaacagg
     1741 tactgggctt tggagtcttg ggaggtggag gtggtgggtg cgaattttac tgctgaattt
     1801 tacacttaat cgcgatctcg cgatcagtgt tttgtgttca cctgtctgtc catctgtctt
     1861 tctgtctgtc agtggcatct cataaaccat cagccctcta ggagtcccag cagttggctt
     1921 aagagactaa tttcaagccc ttgggagttg gaagtcctgc aggtcccctt gtggggagca
     1981 gttcctctag agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt
     2041 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2101 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2161 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2221 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2281 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2341 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2401 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2461 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2521 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2581 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2641 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2701 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2761 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2821 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2881 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2941 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3001 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3061 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3121 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3181 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3241 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3301 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3361 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3421 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3481 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3541 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3601 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3661 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3721 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3781 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3841 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3901 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3961 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4021 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4081 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4141 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4201 atcatgacat taacctataa aaataggcgt atcacgaggc
//
