LOCUS       sequence-367826-        3774 bp    DNA     circular SYN 15-APR-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 Apr 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-709pUC19-tNGFR-P2A-IL10
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1183..1239
                     /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(1617..1633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1617..1633)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1630..1652)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1805..2393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1885..1904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2510..2534)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2634..3425,3426..3494))
                     /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     3257..3276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3495..3599)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3698..3720
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3712..3729
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3713..3729
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctgaccttcg agctcggtac ccggggaaaa attattctaa gagaggtagc ccatcctaaa
       61 aatagctgta atgcagaagt tcatgttcaa ccaatcattt ttgcttacga tgcaaaaatt
      121 gaaaactaag tttattagag aggttagaga aggaggagct ctaagcagaa aaaatcctgt
      181 gccgggaaac cttgattgtg gctttttaat gaatgaagag gcctccctga gcttacaata
      241 taaaaggggg acagagaggt gaaggtctac acatcagggg cttgctcttg caaaacgaaa
      301 ccacaagaca gacttgcaaa agaaagcatg ggtgctggtg caacggggag ggctatggat
      361 ggaccaaggc tccttcttct tttgctcttg ggcgttagtc tgggtggagc taaagaggca
      421 tgccccaccg gactttacac acattccggt gagtgttgca aggcctgcaa cctgggagaa
      481 ggcgtcgctc aaccctgcgg ggcgaatcaa accgtatgcg aaccttgcct cgattctgtt
      541 acattctccg atgtcgtctc tgcgacggaa ccctgcaagc catgcacgga atgtgtcggg
      601 ttgcaaagta tgtcagctcc ttgcgttgaa gcagacgatg cggtgtgtag atgtgcctat
      661 gggtattatc aagacgaaac gacgggacgg tgtgaagcat gtcgagtatg cgaggcgggc
      721 agtggcctcg tcttctcttg tcaggacaag caaaacactg tctgtgaaga atgccctgat
      781 gggacttata gtgatgaggc gaatcacgtc gatccatgtc ttccctgtac tgtctgtgaa
      841 gacactgaga gacaattgcg agagtgtacg cgatgggccg acgcagagtg cgaggagatc
      901 ccaggtcgat ggataaccag gtcaacccct cctgagggat ctgatagtac cgcgccctcc
      961 acccaagaac ccgaagctcc acctgagcaa gacctcatag cgtccacagt agcaggagtt
     1021 gtcacaaccg tcatgggaag ttcacaacca gtagttacca ggggtaccac ggataacctc
     1081 attccggtgt actgcagcat tctcgccgct gtggttgtcg gacttgttgc atacattgcc
     1141 ttcaaacgct ggaatagccg cgccaagcgc tcgggttcgg gtgccaccaa cttcagcctg
     1201 ctgaagcagg ccggcgacgt ggaggagaac cccggccccc acagctcagc actgctctgt
     1261 tgtctggttc tgctcacagg ggtgagggcc agcccaggcc agggcaccca gtctgagaac
     1321 agctgcaccc acttcccagg caacctgcct aacatgcttc gagatctccg agatgccttc
     1381 agcagagtga agactttctt tgtgagtatg attccttcct gtcctttctc tcttcctggg
     1441 actgcctgaa ctagacattc tcctggaact ataagaaccc tcctcctgcg cctccacctc
     1501 catccccaac acctattccc ccaaacttaa attcttaagt cctctagagt cgacctgcag
     1561 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
     1621 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
     1681 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
     1741 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1801 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1921 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1981 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2041 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2101 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2161 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2221 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2281 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2341 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2401 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc
     2461 agcgtaaatg ggtagggggc ttcaaatcgt cctcgtgata ccaattcgga gcctgctttt
     2521 ttgtacaaac ttgttgataa tggcaattca aggatcttca cctagatcct tttaaattaa
     2581 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2641 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2701 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2761 gcaatgatac cgcgagagcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2821 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2881 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2941 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3001 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3061 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3121 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3181 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3241 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3301 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3361 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3421 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3481 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3541 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3601 acatttcccc gaaaagtgcc agatacctga aacaaaaccc atcgtacggc caaggaagtc
     3661 tccaataact gtgatccacc acaagcgcca gggttttccc agtcacgacg ttgtaaaacg
     3721 acggccagtc atgcataatc cgcacgcatc tggaataagg aagtgccatt ccgc
//
