LOCUS Exported 5429 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION A mammalian expression plasmid encoding CD8a with a C-terminus EGFP.. ACCESSION . VERSION . KEYWORDS CD8a-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5429) AUTHORS Madugula V, Lu L TITLE A ternary complex comprising transportin1, Rab8 and the ciliary targeting signal directs proteins to ciliary membranes. JOURNAL J Cell Sci. 2016 Oct 15;129(20):3922-3934. Epub 2016 Sep 15. PUBMED 27633000 REFERENCE 2 (bases 1 to 5429) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5429 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 8..596 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 497..516 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 771..1074 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1075..1278 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1229..1249 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 2079..2088 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2085..2804 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2085..2801 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2130..2151) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2391..2410) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2738..2759 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2927..3048 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2964..2983) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3018..3037 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3055..3510) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3192..3213) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3404..3423 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3537..3641 /gene="bla" /label=AmpR promoter promoter 3643..4000 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3666..3686) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3851..3986 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3913..3932 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4035..4829 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4089..4108) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4699..4718 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5017..5036) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5061..5108 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa 721 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa 781 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata 841 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag 901 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc 961 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta 1021 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg 1081 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt 1141 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca 1201 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag 1261 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag 1321 atctcgagat ggccttacca gtgaccgcct tgctcctgcc gctggccttg ctgctccacg 1381 ccgccaggcc gagccagttc cgggtgtcgc cgctggatcg gacctggaac ctgggcgaga 1441 cagtggagct gaagtgccag gtgctgctgt ccaacccgac gtcgggctgc tcgtggctct 1501 tccagccgcg cggcgccgcc gccagtccca ccttcctcct atacctctcc caaaacaagc 1561 ccaaggcggc cgaggggctg gacacccagc ggttctcggg caagaggttg ggggacacct 1621 tcgtcctcac cctgagcgac ttccgccgag agaacgaggg ctactatttc tgctcggccc 1681 tgagcaactc catcatgtac ttcagccact tcgtgccggt cttcctgcca gcgaagccca 1741 ccacgacgcc agcgccgcga ccaccaacac cggcgcccac catcgcgtcg cagcccctgt 1801 ccctgcgccc agaggcgtgc cggccagcgg cggggggcgc agtgcacacg agggggctgg 1861 acttcgcctg tgatatctac atctgggcgc ccttggccgg gacttgtggg gtccttctcc 1921 tgtcactggt tatcaccctt tactgcaacc acaggaaccg aagacgtgtt tgcaaatgtc 1981 cccggcctgt ggtcaaatcg ggagacaagc ccagcctttc ggcgagatac gtccgaattc 2041 tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg 2101 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc 2161 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga 2221 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga 2281 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca 2341 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca 2401 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc 2461 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact 2521 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact 2581 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga 2641 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt 2701 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga 2761 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta 2821 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca 2881 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc 2941 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 3001 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg 3061 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 3121 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 3181 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 3241 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa 3301 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat 3361 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 3421 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 3481 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg 3541 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 3601 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca 3661 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 3721 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 3781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 3841 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 3901 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 3961 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat 4021 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 4081 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 4141 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 4201 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 4261 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 4321 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 4381 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 4441 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 4501 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 4561 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4621 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4681 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4741 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4801 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 4861 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 4921 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 4981 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc 5041 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc 5101 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag 5161 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc 5221 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg 5281 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 5341 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 5401 agcgtcagac cccgtagaaa agatcaaag //