LOCUS Exported 8487 bp ds-DNA circular SYN 17-MAR-2022 DEFINITION Mouse full length TGF-beta-3 expression. ACCESSION . VERSION . KEYWORDS pD2529-CAG-Flag-mouse TGFb3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8487) AUTHORS Jiang A, Qin Y, Springer TA TITLE Loss of LRRC33-dependent TGFbeta1 activation enhances anti-tumor immunity and checkpoint blockade therapy. JOURNAL Cancer Immunol Res. 2022 Feb 18. pii: 681626. doi: 10.1158/2326-6066.CIR-21-0593. PUBMED 35181792 REFERENCE 2 (bases 1 to 8487) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 17, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8487 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 3..306 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 308..583 /label=chicken beta-actin promoter intron 584..1600 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1524..1546 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1608..1627 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1785..1808 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" polyA_signal 4558..4613 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4612..4631) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" rep_origin complement(5323..5911) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5403..5422) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5997..6806) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 6720..6739 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" promoter complement(6825..6872) /label=EM7 promoter /note="synthetic bacterial promoter " CDS complement(7298..7897) /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(7381..7401) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 7878..7897 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" promoter complement(7907..8406) /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(7955..7974) /label=mPGK-F /note="Mouse PGK promoter, forward primer" primer_bind 8348..8371 /label=MSCV-rev /note="Murine stem cell virus, reverse primer" ORIGIN 1 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 61 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 121 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 181 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 241 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 301 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca 361 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg 421 gggggggcgc gcgccaggcg gggcggggsg gggsgrgggg sggggsgggg sgrggcggrg 481 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg 541 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg 601 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac 661 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg 721 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg 781 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg 841 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg 901 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg 961 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc 1021 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc 1081 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt 1141 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg 1201 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc 1261 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc 1321 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg 1381 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc 1441 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct 1501 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc 1561 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt 1621 tattgtgctg tctcatcatt ttggcaaaga attcggcttg atcgaagccg tctcagggga 1681 gagaccacac ccaagctgtc tagagccgcc accttgtgaa gagcgccgcc accatgcctt 1741 tgctcctcct gttgcccttg ctctgggctg gcgccctcgc ggccgactac aaggacgacg 1801 acgacaagct cgagacttgc accacgttgg acttcggcca catcaagaag aagagggtgg 1861 aagccattag gggacagatc ttgagcaagc tcaggctcac cagcccccct gagccatcgg 1921 tgatgaccca cgtcccctat caggtcctgg cactttacaa cagcacccgg gagttgctgg 1981 aagagatgca cggggagagg gaggaaggct gcactcagga gacctcggag tctgagtact 2041 atgccaaaga gatccataaa ttcgacatga tccagggact ggcggagcac aatgaactgg 2101 ccgtctgccc caaaggaatt acctctaagg tttttcgttt caatgtgtcc tcagtggaga 2161 aaaatggaac caatctgttc cgggcagagt tccgggtctt gcgggtgccc aaccccagct 2221 ccaagcgcac agagcagaga attgagctct tccagatact tcgaccggat gagcacatag 2281 ccaagcagcg ctacataggt ggcaagaatc tgcccacaag gggcaccgct gaatggctgt 2341 ctttcgatgt cactgacact gtgcgcgagt ggctgttgag gagagagtcc aacttgggtc 2401 tggaaatcag catccactgt ccatgtcaca cctttcagcc caatggagac atactggaaa 2461 atgttcatga ggtgatggaa atcaaattca aaggagtgga caatgaagat gaccatggcc 2521 gtggagacct ggggcgtctc aagaagcaaa aggatcacca caacccacac ctgatcctca 2581 tgatgatccc cccacaccga ctggacagcc caggccaggg cagtcagagg aagaagaggg 2641 ccctggacac caattactgc ttccgcaacc tggaggagaa ctgctgtgta cgcccccttt 2701 atattgactt ccggcaggat ctaggctgga aatgggtcca cgaacctaag ggttactatg 2761 ccaacttctg ctcaggccct tgcccatacc tccgcagcgc agacacaacc catagcacgg 2821 tgcttggact atacaacacc ctgaacccag aggcgtctgc ctcgccatgc tgcgtccccc 2881 aggacctgga gcccctgacc atcttgtact atgtgggcag aacccccaag gtggagcagc 2941 tgtccaacat ggtggtgaag tcgtgtaagt gcagctgata agctagcggt tgaggtctct 3001 aaaagcgtct tcctgttctc atcacatcat atcaaggtta tataccatca atattgccac 3061 agatgttact tagcctttta atatttctct aatttagtgt atatgcaatg atagttctct 3121 gatttctgag attgagtttc tcatgtgtaa tgattattta gagtttctct ttcatctgtt 3181 caaatttttg tctagtttta ttttttactg atttgtaaga cttcttttta taatctgcat 3241 attacaattc tctttactgg ggtgttgcaa atattttctg tcattctatg gcctgacttt 3301 tcttaatggt tttttaattt taaaaataag tcttaatatt catgcaatct aattaacaat 3361 cttttctttg tggttaggac tttgagtcat aagaaatttt tctctacact gaagtcatga 3421 tggcatgctt ctatattatt ttctaaaaga tttaaagttt tgccttctcc atttagactt 3481 ataattcact ggaatttttt tgtgtgtatg gtatgacata tgggttccct tttatttttt 3541 acatataaat atatttccct gtttttctaa aaaagaaaaa gatcatcatt ttcccattgt 3601 aaaatgccat atttttttca taggtcactt acatatatca atgggtctgt ttctgagctc 3661 tactctattt tatcagcctc actgtctatc cccacacatc tcatgctttg ctctaaatct 3721 tgatatttag tggaacattc tttcccattt tgttctacaa gaatattttt gttattgtct 3781 ttgggctttc tatatacatt ttgaaatgag gttgacaagt taataacagg cctattgatt 3841 ggaaagtttg tcaacgaatt gtgggtcttt tggggtttgc tgcccctttt acgcaatgtg 3901 gatatcctgc tttaatgcct ttatatgcat gtatacaagc aaaacaggct tttactttct 3961 cgccaactta caaggccttt ctcagtaaac agtatatgac cctttacccc gttgctcggc 4021 aacggcctgg tctgtgccaa gtgtttgctg acgcaacccc cactggttgg ggcttggcca 4081 taggccatca gcgcatgcgt ggaacctttg tgtctcctct gccgatccat actgcggaac 4141 tcctagccgc ttgttttgct cgcagcaggt ctggagcaaa cctcatcggg accgacaatt 4201 ctgtcgtact ctcccgcaag tatacatcgt ttccatggct gctaggctgt gctgccaact 4261 ggatcctgcg cgggacgtcc tttgtttacg tcccgtcggc gctgaatccc gcggacgacc 4321 cctcccgggg ccgcttgggg ctctaccgcc cgcttctccg tctgccgtac cgtccgacca 4381 cggggcgcac ctctctttac gcggactccc cgtctgtgcc ttctcatctg ccggaccgtg 4441 tgcacttcgc ttcacctctg cacgtcgcat ggaggccacc gtgaacgccc accggaacct 4501 gcccaaggtc ttgcataaga ggactcttgg actttcagca atgtcatctc agtggctaat 4561 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa 4621 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg 4681 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt 4741 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt 4801 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct 4861 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc 4921 ctcttctctt atggagatct ttgcattttt agacatttag aagcctatat cttgttacag 4981 aattggaatt acacaaaaat tctaccatat tttgaaagct taggttgttc tgaaaaaaac 5041 aatatattgt tttcctgggt aaactaaaag tcccctcgag gaaaggcccc taaagtgaaa 5101 cagtgcaaaa cgttcaaaaa ctgtctggca atacaagttc cactttgacc aaaacggctg 5161 gcagtaaaag ggttaagaag actgtcagcc ttgagcggta tcagctcact caaaggcggt 5221 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 5281 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 5341 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 5401 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 5461 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 5521 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 5581 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 5641 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 5701 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg gctaactacg gctacactag 5761 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 5821 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 5881 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 5941 tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag ggattttggt catgagttag 6001 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca 6061 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg 6121 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt 6181 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa 6241 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca 6301 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc 6361 tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgagtgc 6421 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct 6481 tctaatacct ggaacgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca 6541 ggagtacgga taaaatgctt gatggtcgga agtggcataa attccgtcag ccagtttagt 6601 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac 6661 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta 6721 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc 6781 gacgtttccc gttggatatg gctcattttt tacttcctca ccttgtcgta ttatactatg 6841 ccgatatact atgccgatga ttaattgtcg acactgcggg ggctctgtgt ggtaagcagg 6901 tcttaacctt tttactgcca atgacgcatg ggatacgtcg tggcagtaaa agggcttaaa 6961 tgccaacgac gcgtcccata cgttgttggc attttaattc ttctctctgc agcggcagca 7021 tgtgccgccg ctgcagagag tttctagcga tgacagcccc tctgggcaac gagccggggg 7081 ggctgtcttt ctttatgttt taaatgcact gacctcccac attccctttt tagtaaaata 7141 ttcagaaata atttaaatac atcattgcaa tgaaaataaa tgttttttat taggcagaat 7201 ccagatgctc aaggcccttc ataatatccc ccagtttagt agttggactt agggaacaaa 7261 ggaaccttta atagaaattg gacagcaaga aagcgagtca ggcaccgggc ttgcgggtca 7321 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc 7381 gctcgtagaa ggggaggttg cggggcgcgg atgtctccag gaaggcgggc accccggcgc 7441 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt 7501 cgggcgacac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca 7561 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg 7621 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca 7681 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga 7741 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg 7801 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc 7861 gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt ggcggacgaa aggcccggag 7921 atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc 7981 ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc ccgcccccgg 8041 acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc 8101 tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag 8161 tgagtcgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga 8221 acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg 8281 gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc 8341 aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc 8401 ctacccggta gagaaacttg atctgtcgcc gcaattcaaa cttcgtgagg ctccggtgcc 8461 cgtcagtgac ctgctatact ctggaga //