LOCUS Exported 7540 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCAGGS-ChR2-Venus SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7540) AUTHORS Petreanu L, Huber D, Sobczyk A, Svoboda K TITLE Channelrhodopsin-2-assisted circuit mapping of long-range callosal projections. JOURNAL Nat Neurosci. 2007 May . 10(5):663-8. PUBMED 17435752 REFERENCE 2 (bases 1 to 7540) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7540 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..661 /label=chicken beta-actin promoter intron 662..1670 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1594..1616 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" CDS 1758..2684 /codon_start=1 /product="N-terminal fragment of channelrhodopsin-2 from the alga Chlamydomonas reinhardtii (Nagel et al., 2003)" /label=ChR2 /note="blue light-activated cation channel" /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI RKTTKLNIGGTEIEVETLVEDEAEAGAV" regulatory 2712..2721 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2718..3437 /codon_start=1 /product="Venus YFP with monomerizing A206K mutation (Nagai et al., 2002; Kremers et al., 2006)" /label=mVenus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2763..2784) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3024..3043) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3371..3392 /label=EGFP-C /note="EGFP, forward primer" intron 3463..3596 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 3579..3598 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 3605..4193 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3658..3678) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4076..4087) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4220..4237) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4226..4337 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4564..4583) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4629..4684 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4683..4702) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(5043..5059) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5043..5059) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5056..5078) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5067..5083 /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(5091..5121) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5136..5157 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5216..5411 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5262..5397 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5324..5343 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5417..5551 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5454..5473) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5508..5527 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5619..5636) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5790..6378) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5870..5889) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6549..7409) /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 7172..7191 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7410..7514) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg 541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag 841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct 1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc 1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt 1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg 1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg 1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca 1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcggtacc gcgggcccgg 1741 gatccaccgg tgctagcatg gattatggag gcgccctgag tgccgttggg cgcgagctgc 1801 tatttgtaac gaacccagta gtcgtcaatg gctctgtact tgtgcctgag gaccagtgtt 1861 actgcgcggg ctggattgag tcgcgtggca caaacggtgc ccaaacggcg tcgaacgtgc 1921 tgcaatggct tgctgctggc ttctccatcc tactgcttat gttttacgcc taccaaacat 1981 ggaagtcaac ctgcggctgg gaggagatct atgtgtgcgc tatcgagatg gtcaaggtga 2041 ttctcgagtt cttcttcgag tttaagaacc cgtccatgct gtatctagcc acaggccacc 2101 gcgtccagtg gttgcgttac gccgagtggc ttctcacctg cccggtcatt ctcattcacc 2161 tgtcaaacct gacgggcttg tccaacgact acagcaggcg caccatgggt ctgcttgtgt 2221 ctgatattgg cacaattgtg tggggcgcca cttccgccat ggccaccgga tacgtcaagg 2281 tcatcttctt ctgcctgggt ctgtgttatg gtgctaacac gttctttcac gctgccaagg 2341 cctacatcga gggttaccac accgtgccga agggccggtg tcgccaggtg gtgactggca 2401 tggcttggct cttcttcgta tcatggggta tgttccccat cctgttcatc ctcggccccg 2461 agggcttcgg cgtcctgagc gtgtacggct ccaccgtcgg ccacaccatc attgacctga 2521 tgtcgaagaa ctgctggggt ctgctcggcc actacctgcg cgtgctgatc cacgagcata 2581 tcctcatcca cggcgacatt cgcaagacca ccaaattgaa cattggtggc actgagattg 2641 aggtcgagac gctggtggag gacgaggccg aggctggcgc ggtcaacaag ggcaccggca 2701 agccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca 2761 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg 2821 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc 2881 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct 2941 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 3001 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 3061 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 3121 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg 3181 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg 3241 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 3301 tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga 3361 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 3421 acgagctgta caagtaaagc ggccccggac tcgaggccgc aggtaagtat caaggttaca 3481 agacaggttt aaggagacca catagaaact gggcttgtcg agacagagaa gactcttgcg 3541 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt 3601 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 3661 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 3721 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 3781 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 3841 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 3901 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 3961 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct 4021 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 4081 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 4141 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat 4201 tcgagctcgg tacgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 4261 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 4321 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 4381 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggccagcttt tgttcccttt 4441 agggttaatt gcgcgcttgg cgctaatcat ggtcatagct gtttccactc ctcaggtgca 4501 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg 4561 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac 4621 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct 4681 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt 4741 ttagagtttg gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag 4801 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga 4861 cttgaggtta gatttttttt tatattttgt ttgtgttatt tttttcttta acatccctaa 4921 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca 4981 tagctgtccc tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca 5041 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5101 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5161 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc 5221 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 5281 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 5341 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 5401 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag 5461 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 5521 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg 5581 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 5641 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 5701 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 5761 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 5821 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 5881 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 5941 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg 6001 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 6061 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 6121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 6181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 6241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 6301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 6361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 6421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 6481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6601 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6661 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 6721 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 6781 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 6841 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6901 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 6961 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 7021 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 7081 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 7141 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 7201 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 7261 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 7321 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 7381 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 7441 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7501 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg //