LOCUS pIG-830_HA-GD2-28z_CAR_tNGFR_Retroviral 8647 bp DNA circular SYN 14-APR-2025 DEFINITION This plasmid can be used to generate retrovirus.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8647) AUTHORS Blaeschke F, Chen YY, Apathy R, Daniel B, Chen AY, Chen PA, Sandor K, Zhang W, Li Z, Mowery CT, Yamamoto TN, Nyberg WA, To A, Yu R, Bueno R, Kim MC, Schmidt R, Goodman DB, Feuchtinger T, Eyquem J, Jimmie Ye C, Carnevale J, Satpathy AT, Shifrut E, Roth TL, Marson A TITLE Modular pooled discovery of synthetic knockin sequences to program durable cell therapies. JOURNAL Cell. 2023 Sep 14;186(19):4216-4234.e33. doi: 10.1016/j.cell.2023.08.013. PUBMED 37714135 REFERENCE 2 (bases 1 to 8647) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1 //gt12365.cn COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.cell.2023.08"; journalName: "Cell"; date: "2023-09-14- 14"; volume: "186"; issue: "19"; pages: "4216-4234.e33" COMMENT Sequence Label: pIG-830_HA-GD2-28z_CAR_tNGFR_Retroviral FEATURES Location/Qualifiers source 1..8647 /mol_type="other DNA" /organism="synthetic DNA construct" CDS complement(join(171..962,963..1031)) /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 794..813 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1032..1136) /gene="bla" /label=AmpR promoter primer_bind 1204..1222 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(1260..1282) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 1382..1401 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1595..1617 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1745..1764 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 1964..2480 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" primer_bind 3320..3342 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 3358..3374 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind 3712..3728 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" regulatory 3781..3790 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 4681..4734 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" LTR 6995..7509 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 7681..7697 /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(7705..7711,7712..7729,7730..7735)) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7750..7771 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7888..7905) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8059..8647) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8139..8158) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 1141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat 1201 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac 1261 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc 1321 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat 1381 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga 1441 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg 1501 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct 1561 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg 1621 gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg 1681 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc 1741 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg 1801 ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg 1861 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat 1921 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct 1981 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact 2041 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac 2101 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag 2161 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag 2221 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt 2281 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc 2341 agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt 2401 tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc 2461 cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac 2521 caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct 2581 ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact 2641 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc 2701 ctgggagacg tcccagggac ttcgggggcc gtttttgtgg cccgacctga gtcctaaaat 2761 cccgatcgtt taggactctt tggtgcaccc cccttagagg agggatatgt ggttctggta 2821 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttgggac 2881 cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact 2941 gtgtttctgt atttgtctga aaatatgggc ccgggctagc ctgttaccac tcccttaagt 3001 ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc 3061 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg 3121 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca 3181 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct 3241 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct 3301 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt 3361 tatccagccc tcactccttc tctaggcgcc cccatatggc catatgagat cttatatggg 3421 gcacccccgc cccttgtaaa cttccctgac cctgacatga caagagttac taacagcccc 3481 tctctccaag ctcacttaca ggctctctac ttagtccagc acgaagtctg gagacctctg 3541 gcggcagcct accaagaaca actggaccga ccggtggtac ctcaccctta ccgagtcggc 3601 gacacagtgt gggtccgccg acaccagact aagaacctag aacctcgctg gaaaggacct 3661 tacacagtcc tgctgaccac ccccaccgcc ctcaaagtag acggcatcgc agcttggata 3721 cacgccgccc acgtgaaggc tgccgacccc gggggtggac catcctctag ccctcgagcc 3781 gccaccatgg gcgctggtgc aactggacgg gccatggatg gaccacggct tttgctcctg 3841 ttgttgttgg gcgtcagcct tggaggggct aaagaggcct gcccgaccgg tttgtacaca 3901 cactccggcg agtgttgtaa ggcgtgtaat cttggcgaag gggtcgccca accgtgtggc 3961 gccaaccaga cggtttgtga gccttgcttg gatagcgtca cgttttccga cgtcgtttca 4021 gccaccgaac catgcaaacc ttgtacggaa tgcgtgggtc tgcagagcat gagcgcaccg 4081 tgcgtagagg cggacgatgc ggtatgccga tgtgcctacg gctactacca agacgagacg 4141 acagggaggt gtgaggcatg cagagtatgt gaggccgggt ctggactcgt attttcctgt 4201 caagacaaac aaaacacagt atgtgaggag tgcccggatg ggacttattc cgatgaagcg 4261 aatcatgtgg acccgtgctt gccatgtacg gtctgtgagg ataccgaaag acagctccga 4321 gaatgcaccc gctgggctga cgcggaatgc gaagaaatcc caggtcgatg gatcacccgc 4381 tcaacccccc ccgaagggtc agattctacg gcgccctcta ctcaggagcc agaagctcca 4441 cccgaacaag acctgatcgc ttctacggtg gcaggtgtcg ttacaacagt catgggttcc 4501 tcacaacctg tggttacgcg ggggacaact gacaatttga ttccagtata ttgtagcatt 4561 ctggccgcgg tcgtggtagg gcttgtcgcg tatatagcgt tcaaaaggtg gaacagcagg 4621 gctaagagag gttcaggatc tggggggggc tcgggaggtg gaaccggtgc tggaagtggt 4681 gagggcaggg gatctctcct tacttgtggc gacgtggagg agaaccccgg ccccatggaa 4741 ttcggcctga gctggctgtt cctggtggcc atcctgaagg gcgtgcagtg cagcagagac 4801 atcctgctga cccagacccc tctgagcctg cctgtgtctc tgggcgatca ggccagcatc 4861 agctgcagat ccagccagag cctggtgcac cggaacggca acacctacct gcactggtat 4921 ctgcagaagc ccggccagag ccccaagctg ctgatccaca aggtgtccaa ccggttcagc 4981 ggcgtgcccg acagattttc tggcagcggc tccggcaccg acttcaccct gaagatcagc 5041 cgggtggaag ccgaggacct gggcgtgtac ttctgcagcc agtccaccca cgtgcccccc 5101 ctgacatttg gcgccggaac aaagctggaa ctgaaggggg gaggcggatc tggcggcgga 5161 ggaagtggcg gagggggatc tgaagtgaag ctgcagcagt ccggccccag cctggtggaa 5221 cctggcgcct ctgtgatgat ctcctgcaag gccagcggca gctccttcac cggctacaac 5281 atgaactggg tgcgccagaa catcggcaag agcctggaat ggatcggcgc catcgacccc 5341 tactacggcg gcaccagcta caaccagaag ttcaagggca gagccaccct gaccgtggac 5401 aagagcagca gcaccgccta catgcacctg aagtccctga ccagcgagga cagcgccgtg 5461 tactactgcg tgtccggcat gaagtactgg ggccagggca caagcgtgac cgtgtctagc 5521 gccaagacca ccccccctag cgtgtacgga agagtgacag tgtcctctgc cgagcccaag 5581 agctgcgaca agacccacac ctgtccccct tgtcctgccc ctcctgttgc aggcccttcc 5641 gtgttcctgt tccccccaaa gcccaaggac acactgatga tcagcagaac ccccgaagtg 5701 acctgcgtgg tggtggacgt gtcccacgag gacccagaag tgaagttcaa ttggtacgtg 5761 gacggcgtgg aagtgcacaa cgccaagaca aagcccagag aggaacagta ccaaagcacc 5821 taccgggtgg tgtccgtgct gaccgtgctg catcaggatt ggctgaacgg caaagagtac 5881 aagtgcaaag tgtccaacaa ggccctgcct gcccccatcg agaaaaccat cagcaaggcc 5941 aagggccagc cccgcgaacc ccaggtgtac acactgcccc ctagcaggga cgagctgacc 6001 aagaaccagg tgtccctgac atgcctcgtg aagggcttct acccctccga tatcgccgtg 6061 gaatgggaga gcaacggcca gcccgagaac aactacaaga caacccctcc cgtgctggac 6121 agcgacggct cattcttcct gtacagcaag ctgacagtgg ataagtcccg gtggcagcag 6181 ggcaacgtgt tcagctgctc cgtgatgcac gaggccctgc acaaccacta cacccagaaa 6241 agcctgtccc tgagccccgg caagaaggac cccaaagcgg ccgcaattga agttatgtat 6301 cctcctcctt acctagacaa tgagaagagc aatggaacca ttatccatgt gaaagggaaa 6361 cacctttgtc caagtcccct atttcccgga ccttctaagc ccttttgggt gctggtggtg 6421 gttgggggag tcctggcttg ctatagcttg ctagtaacag tggcctttat tattttctgg 6481 gtgaggagta agaggagcag gctcctgcac agtgactaca tgaacatgac tccccgccgc 6541 cccgggccca cccgcaagca ttaccagccc tatgccccac cacgcgactt cgcagcctat 6601 cgctccagag tgaagttcag caggagcgca gacgcccccg cgtaccagca gggccagaac 6661 cagctctata acgagctcaa tctaggacga agagaggagt acgatgtttt ggacaagaga 6721 cgtggccggg accctgagat ggggggaaag ccgagaagga agaaccctca ggaaggcctg 6781 tacaatgaac tgcagaaaga taagatggcg gaggcctaca gtgagattgg gatgaaaggc 6841 gagcgccgga ggggcaaggg gcacgatggc ctttaccagg gtctcagtac agccaccaag 6901 gacacctacg acgcccttca catgcaggcc ctgccccctc gctaaggatc cgataaaata 6961 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg 7021 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag 7081 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg 7141 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc 7201 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat 7261 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt 7321 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga 7381 tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact 7441 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg 7501 gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga 7561 atattaagta atcctgactc aattagccac tgttttgaat ccacatactc caatactcct 7621 gaaatccatc gatggagttc attatggaca gcgcagaaag agctggggag aattgtgaaa 7681 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 7741 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 7801 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 7861 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 7921 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 7981 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 8041 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 8101 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 8161 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 8221 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 8281 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 8341 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 8401 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 8461 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 8521 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 8581 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 8641 atctcaa //