LOCUS Exported 3774 bp ds-DNA circular SYN 26-JUL-2022 DEFINITION Knockin of truncated NGFR to target gene. ACCESSION . VERSION . KEYWORDS pIG-720pUC19-tNGFR-P2A-ZAP70 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3774) AUTHORS Shy BR, Vykunta V, Ha A, Roth TL, Talbot A, Nguyen DN, Chen YY, Blaeschke F, Vedova S, Mamedov MR, Chung J, Li H, Wolf J, Martin TG, Ye L, Eyquem J, Esensten JH, Alexander M TITLE Hybrid ssDNA repair templates enable high yield genome engineering in primary cells for disease modeling and cell therapy manufacturing JOURNAL bioRxiv 2021.09.02.458799 REFERENCE 2 (bases 1 to 3774) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 26, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3774 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1176..1232 /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(1610..1626) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1610..1626) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1623..1645) /label=M13/pUC Reverse /note="In lacZ gene" rep_origin complement(1798..2386) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1878..1897) /label=pBR322ori-F /note="pBR322 origin, forward primer" protein_bind complement(2503..2527) /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS complement(2627..3487) /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 3250..3269 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3488..3592) /gene="bla" /label=AmpR promoter primer_bind 3691..3713 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3705..3722 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3706..3722 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 tcgagctcgg tacccgggga tgggtacagg tgtgggtgct tggggaccag ttggcgtgct 61 tgggcgtcca cataatgtct ctggaaagtc agatgggggt ttgggacttc tcgatctgtg 121 cccagcaggc tgcggcttct ctccaggttg actctggcac agagcaggct ctgccccctt 181 ggcgagctca gtctgcggca ctgatgccct ccacttggcg tctctcgcgc cgtctttggg 241 cccaacgcac caggttcagg aaggccctga cgtgcctccg accctctgtg aacccgcagg 301 tttcgcgagg cccaggggcg atgggtgctg gtgcaacggg gagggctatg gatggaccaa 361 ggctccttct tcttttgctc ttgggcgtta gtctgggtgg agctaaagag gcatgcccca 421 ccggacttta cacacattcc ggtgagtgtt gcaaggcctg caacctggga gaaggcgtcg 481 ctcaaccctg cggggcgaat caaaccgtat gcgaaccttg cctcgattct gttacattct 541 ccgatgtcgt ctctgcgacg gaaccctgca agccatgcac ggaatgtgtc gggttgcaaa 601 gtatgtcagc tccttgcgtt gaagcagacg atgcggtgtg tagatgtgcc tatgggtatt 661 atcaagacga aacgacggga cggtgtgaag catgtcgagt atgcgaggcg ggcagtggcc 721 tcgtcttctc ttgtcaggac aagcaaaaca ctgtctgtga agaatgccct gatgggactt 781 atagtgatga ggcgaatcac gtcgatccat gtcttccctg tactgtctgt gaagacactg 841 agagacaatt gcgagagtgt acgcgatggg ccgacgcaga gtgcgaggag atcccaggtc 901 gatggataac caggtcaacc cctcctgagg gatctgatag taccgcgccc tccacccaag 961 aacccgaagc tccacctgag caagacctca tagcgtccac agtagcagga gttgtcacaa 1021 ccgtcatggg aagttcacaa ccagtagtta ccaggggtac cacggataac ctcattccgg 1081 tgtactgcag cattctcgcc gctgtggttg tcggacttgt tgcatacatt gccttcaaac 1141 gctggaatag ccgcgccaag cgctcgggtt cgggtgccac caacttcagc ctgctgaagc 1201 aggccggcga cgtggaggag aaccccggcc ccccagaccc cgccgcccat ctgccctttt 1261 tttatgggag catctcgcgt gccgaggccg aggagcacct gaagctggcg ggcatggcgg 1321 acgggctctt cctgctgcgc cagtgcctgc gctcgctggg cggctatgtg ctgtcgctcg 1381 tgcacgatgt gcgcttccac cactttccca tcgagcgcca gctcaacggc acctacgcca 1441 ttgccggcgg caaagcgcac tgtggaccgg cagagctctg cgagttctac tcgcgcgacc 1501 ccgacgggct gccctgcaac ctgcgcaagc cgtcctctag agtcgacctg cagaggctag 1561 gtggaggctc agtgatgata agtctgcgat ggtggatgca tgtgtcatgg tcatagctgt 1621 ttcctgtgtg aaattgttat ccgctcagag ggcacaatcc tattccgcgc tatccgacaa 1681 tctccaagac attaggtgga gttcagttcg gcgtatggca tatgtcgctg gaaagaacat 1741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 1801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 1861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 1921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 1981 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 2041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 2101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 2161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 2221 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 2281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 2341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 2401 cttttctacg gggtctgacg ctctattcaa caaagccgcc gtcccgtcaa gtcagcgtaa 2461 atgggtaggg ggcttcaaat cgtcctcgtg ataccaattc ggagcctgct tttttgtaca 2521 aacttgttga taatggcaat tcaaggatct tcacctagat ccttttaaat taaaaatgaa 2581 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 2641 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 2701 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 2761 taccgcgaga gccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 2821 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 2881 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 2941 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 3001 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 3061 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 3121 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 3181 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 3241 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 3301 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 3361 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 3421 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 3481 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 3541 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 3601 cccgaaaagt gccagatacc tgaaacaaaa cccatcgtac ggccaaggaa gtctccaata 3661 actgtgatcc accacaagcg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 3721 gtcatgcata atccgcacgc atctggaata aggaagtgcc attccgcctg acct //