LOCUS Exported 5436 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Constitutive CAG-driven expression of mGreenLantern in animals using AAV. ACCESSION . VERSION . KEYWORDS pAAV-CAG-mGreenLantern SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5436) AUTHORS Campbell BC, Nabel EM, Murdock MH, Lao-Peregrin C, Tsoulfas P, Blackmore MG, Lee FS, Liston C, Morishita H, Petsko GA TITLE mGreenLantern: a bright monomeric fluorescent protein with rapid expression and cell filling properties for neuronal imaging. JOURNAL Proc Natl Acad Sci U S A. 2020 Dec 1;117(48):30710-30721. doi: 10.1073/pnas.2000942117. Epub 2020 Nov 18. PUBMED 33208539 REFERENCE 2 (bases 1 to 5436) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5436 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 418..692 /label=chicken beta-actin promoter primer_bind 971..993 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1055..1074 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1107..1116 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1113..1832 /codon_start=1 /product="monomeric GFP derivative that displays exceptionally high brightness in cells (Campbell et al., 2020)" /label=mGreenLantern /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK ERVTAAGITHDMDELYK" primer_bind complement(1158..1179) /label=EGFP-N /note="EGFP, reverse primer" misc_feature 1857..2445 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1910..1930) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2328..2339) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2488..2609 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2499..2518) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2553..2572 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 2771..2900 /label=AAV2 ITR primer_bind complement(2966..2983) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3137..3725) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3217..3236) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3896..4756) /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 4519..4538 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4757..4861) /gene="bla" /label=AmpR promoter rep_origin complement(4887..5342) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5024..5045) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5236..5255 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 5414..107 /label=AAV2 ITR ORIGIN 1 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga 61 gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt agttaatgat 121 taacccgcca tgctacttat ctacgtagcc atgctctagg aagagtacca ttgacgtcaa 181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg 421 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt 481 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggcgcg 541 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg 601 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc 661 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg 721 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc 781 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat 841 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt 901 tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg 961 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat 1021 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct 1081 catcattttg gcaaagaatt ggatccgcca ccatggtgag caagggcgag gagctgttca 1141 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 1201 tccgcggcga gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca 1261 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttaggc tacggcgtgg 1321 cctgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 1381 ccgaaggcta cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc 1441 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgtgctg aagggcatcg 1501 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca 1561 aggtctatat cacggccgac aagcagaaga acggcatcaa ggctaacttc aagacccgcc 1621 acaacgttga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 1681 gcgacggccc cgtgctgctg cccgacaacc actacctgag ccatcagtcc aaactgagca 1741 aagaccccaa cgagaagcgc gatcacatgg tcctgaagga gagggtgacc gccgccggga 1801 ttacacatga catggacgag ctgtacaagt aagaattcga tatcaagctt atcgataatc 1861 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 1921 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 1981 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 2041 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 2101 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 2161 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 2221 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct 2281 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 2341 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 2401 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac 2461 ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa 2521 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg 2581 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc 2641 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta 2701 aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc 2761 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg 2821 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca 2881 gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 2941 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 3001 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 3061 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 3121 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 3181 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 3241 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 3301 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 3361 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 3421 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 3481 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 3541 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 3601 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 3661 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 3721 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 3781 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 3841 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 3901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 3961 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 4021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 4081 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 4141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 4201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 4261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 4321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 4381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 4441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 4501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 4561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 4621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 4681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 4741 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 4801 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 4861 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc 4921 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 4981 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 5041 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 5101 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 5161 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 5221 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 5281 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 5341 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 5401 gctattacgc cagctgcgcg ctcgctcgct cactga //