LOCUS Exported 5520 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression vector for expression of DsRed. ACCESSION . VERSION . KEYWORDS pCAG-DsRed SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5520) 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 5520) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5520 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 6..385 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 387..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" regulatory 1772..1781 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1778..2458 /codon_start=1 /product="wild-type DsRed" /label=DsRed1 /note="mammalian codon-optimized" /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH HLFL" primer_bind complement(1961..1981) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 2179..2198 /label=mCherry-F /note="mCherry, forward primer" primer_bind 2373..2396 /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(2543..2562) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2608..2663 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2662..2681) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3024..3040) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3024..3040) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3037..3059) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3048..3064 /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(3072..3102) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3117..3138 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3197..3392 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3243..3378 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3305..3324 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 3398..3532 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3435..3454) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3489..3508 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3600..3617) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3771..4359) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3851..3870) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4530..5390) /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 5153..5172 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5391..5495) /gene="bla" /label=AmpR promoter ORIGIN 1 gggtcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca 61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc 121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat 181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt 241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc 301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta 361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc 421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc 481 agcgatgggg gcgggggggg gggggncgcg cgccaggcgg 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 ttctgcagtc 1741 gacggtaccg cgggcccggg atccaccggt cgccaccatg gtgcgctcct ccaagaacgt 1801 catcaaggag ttcatgcgct tcaaggtgcg catggagggc accgtgaacg gccacgagtt 1861 cgagatcgag ggcgagggcg agggccgccc ctacgagggc cacaacaccg tgaagctgaa 1921 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta 1981 cggctccaag gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt 2041 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt 2101 gacccaggac tcctccctgc aggacggctg cttcatctac aaggtgaagt tcatcggcgt 2161 gaacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac 2221 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccacaagg ccctgaagct 2281 gaaggacggc ggccactacc tggtggagtt caagtccatc tacatggcca agaagcccgt 2341 gcagctgccc ggctactact acgtggactc caagctggac atcacctccc acaacgagga 2401 ctacaccatc gtggagcagt acgagcgcac cgagggccgc caccacctgt tcctgtagcg 2461 gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc 2521 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga 2581 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt 2641 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac 2701 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga 2761 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct 2821 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat 2881 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc 2941 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc 3001 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 3061 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 3121 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 3181 tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3241 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3301 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag 3361 gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa 3421 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 3481 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat 3541 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 3601 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 3661 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 3721 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 3781 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 3841 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 3901 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 3961 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 4021 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 4081 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 4141 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 4201 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 4261 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 4321 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 4381 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 4441 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 4501 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 4561 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 4621 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 4681 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 4741 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 4801 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 4861 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 4921 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 4981 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 5041 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 5101 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 5161 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 5221 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 5281 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 5341 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 5401 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 5461 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct //