LOCUS Exported 6124 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression vector for cloning and expressing your gene on a bicistronic mRNA also encoding IRES-GFP. ACCESSION . VERSION . KEYWORDS pCAGIG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6124) AUTHORS Matsuda T, Cepko CL TITLE Electroporation and RNA interference in the rodent retina in vivo and in vitro. JOURNAL Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22. PUBMED 14603031 REFERENCE 2 (bases 1 to 6124) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6124 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 5..384 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..662 /label=chicken beta-actin promoter intron 663..1679 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1603..1625 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1687..1706 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" misc_feature 1776..2362 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1943..1960) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2170..2189 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2363..3082 /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 2363..3079 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2408..2429) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2669..2688) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3016..3037 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3146..3165) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3211..3266 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3265..3284) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3627..3643) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3627..3643) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3640..3662) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3651..3667 /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(3675..3705) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3720..3741 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3800..3995 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3846..3981 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3908..3927 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4001..4135 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4038..4057) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4092..4111 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4203..4220) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4374..4962) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4454..4473) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5133..5993) /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 5756..5775 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5994..6098) /gene="bla" /label=AmpR promoter ORIGIN 1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat 61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg 121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggggcgc gcncaggcgg ggcggggcgg ggcgaggggc 541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt 601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg 661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc 721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc 781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg 841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt 901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc 961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg 1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg 1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc 1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc 1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg 1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg 1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg 1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca 1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg 1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc 1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg 1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc 1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgagg 1741 atatcgcggc cgctacgtaa attccgcccc ccccccccct ctccctcccc cccccctaac 1801 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc 1861 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg 1921 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg 1981 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc 2041 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa 2101 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa 2161 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta 2221 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg 2281 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca 2341 cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt 2401 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 2461 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 2521 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 2581 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 2641 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 2701 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 2761 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 2821 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 2881 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 2941 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 3001 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 3061 catggacgag ctgtacaagt aaagcggccg gccgccagca cagtggtcga tcgaccaatg 3121 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca 3181 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag 3241 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa 3301 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca 3361 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt 3421 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt 3481 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc 3541 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc 3601 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 3661 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 3721 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 3781 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc 3841 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3901 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 3961 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta 4021 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4081 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg 4141 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 4201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 4261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 4321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 4381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 4441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 4501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 4561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 4621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 4681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 4741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 4801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 4861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 4921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 4981 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 5041 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 5101 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 5161 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 5221 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 5281 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 5341 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 5401 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 5461 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 5521 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 5581 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 5641 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 5701 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 5761 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 5821 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 5881 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 5941 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 6001 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 6061 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 6121 cctg //