LOCUS Exported 4803 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Longitudinal monitoring of Gaussia and Nano luciferase activities to simultaneously assess ER calcium homeostasis and ER stress in vivo. ACCESSION . VERSION . KEYWORDS pOTTC841 - pAAV minP secNLuc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4803) AUTHORS Wires ES, Henderson MJ, Yan X, Back S, Trychta KA, Lutrey MH, Harvey BK TITLE Longitudinal monitoring of Gaussia and Nano luciferase activities to concurrently assess ER calcium homeostasis and ER stress in vivo. JOURNAL PLoS One. 2017 Apr 12;12(4):e0175481. doi: 10.1371/journal.pone.0175481. eCollection 2017. PUBMED 28403212 REFERENCE 2 (bases 1 to 4803) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4803 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR CDS 368..880 /codon_start=1 /product="NanoLuc(R) luciferase" /label=Nluc /note="human codon-optimized" /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR LCERILA" misc_feature 930..1518 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(983..1003) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1401..1412) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 1550..2026 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(1749..1768) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 2066..2206 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2066..2195 /label=AAV2 ITR rep_origin 2281..2736 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2368..2387) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2578..2599 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2753..2772) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2872..2894 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2932..2950) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3018..3122 /gene="bla" /label=AmpR promoter CDS 3123..3983 /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 complement(3341..3360) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4154..4742 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4643..4662 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtcactc attcataaaa cgcttgttat 181 aaaagcagtg gctgcggcgc ctcgtactcc aaccgcatct gcagcgagca actgagaagc 241 caagactgag ccggcggccg gtactgttgg taaagccacc atgaactcct tctccacaag 301 cgccttcggt ccagttgcct tctccctggg cctgctcctg gtgttgcctg ctgccttccc 361 tgccccagtc ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa 421 cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc 481 cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca 541 tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt 601 taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact 661 ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat 721 cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat 781 tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg 841 agtgaccggc tggcggctgt gcgaacgcat tctggcgtaa ggccgcgact ctagagtggc 901 gcgccgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa 961 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 1021 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 1081 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 1141 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 1201 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 1261 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 1321 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga 1381 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 1441 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 1501 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc 1561 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 1621 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 1681 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc 1741 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat 1801 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca 1861 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat 1921 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa 1981 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt 2041 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 2101 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 2161 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 2221 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 2281 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 2341 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 2401 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 2461 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 2521 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 2581 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat 2641 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 2701 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 2761 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 2821 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 2881 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 2941 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 3001 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 3061 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 3121 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 3181 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 3241 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 3301 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 3361 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 3421 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 3481 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 3541 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 3601 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 3661 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 3721 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 3781 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 3841 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 3901 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 3961 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 4021 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 4081 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 4141 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 4201 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 4261 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt 4321 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 4381 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 4441 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 4501 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 4561 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 4621 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 4681 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 4741 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 4801 tgt //