LOCUS Exported 6227 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION FLiCRE protease for neuronal expression. ACCESSION . VERSION . KEYWORDS mtagBFP2-CaM-uTEVp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6227) AUTHORS Kim CK, Sanchez MI, Hoerbelt P, Fenno LE, Malenka RC, Deisseroth K, Ting AY TITLE A Molecular Calcium Integrator Reveals a Striatal Cell Type Driving Aversion. JOURNAL Cell. 2020 Dec 23;183(7):2003-2019.e16. doi: 10.1016/j.cell.2020.11.015. Epub 2020 Dec 11. PUBMED 33308478 REFERENCE 2 (bases 1 to 6227) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6227 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 181..628 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 642..651 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 648..1358 /codon_start=1 /product="enhanced monomeric blue fluorescent protein (Subach et al., 2011)" /label=mTagBFP2 /note="mammalian codon-optimized" /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV ARYCDLPSKLGHKLN" CDS 1395..1838 /codon_start=1 /product="calcium-binding protein" /label=calmodulin /translation="MDQLTEEQIAEFKEAFSLLDKDGDGTITTKELGTGMRSLGQNPTE AELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGNGYISAAE LRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK" CDS 1839..1880 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 2619..3207 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2672..2692) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3090..3101) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3249..3266) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3255..3462 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3491..3631 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3491..3620 /label=AAV2 ITR rep_origin 3705..4160 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3792..3811) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4002..4023 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4177..4196) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4296..4318 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4356..4374) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4442..4546 /gene="bla" /label=AmpR promoter CDS 4547..5407 /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(4765..4784) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5578..6166 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6067..6086 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg 181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg 601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg gtgtctaagg 661 gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca 721 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca 781 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta 841 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca 901 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg 961 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga 1021 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg 1081 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg 1141 ccctgaagct cgtgggcggg agccatctga tcgcaaacgc caagaccaca tatagatcca 1201 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa 1261 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat 1321 actgcgacct ccctagcaaa ctggggcaca agcttaatga ccttggatct ggcacaggcg 1381 gatccggttc aggcatggac caactgactg aagagcagat cgcagaattt aaagaggctt 1441 tctccctact ggacaaggac ggggatggga caataacaac caaggagctg gggacgggga 1501 tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag 1561 atgccgacgg tgacggcaca atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa 1621 tgaaatacac agacagtgaa gaagaaatta gagaagcgtt ccgtgtgttt gataaggatg 1681 gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt 1741 taacagatga agaggttgat gaaatgatca gggaagcaga catcgatggg gatggtcagg 1801 taaactacga agagtttgta caaatgatga cagcgaaggg caagccgatc cctaaccctc 1861 tgctgggcct ggacagcact ggtggaagtg gatcaggcag cggtggatct tacggaagcc 1921 atgtggatta cgccggagaa tccttgttta agggaccacg tgattacaac ccgatatcga 1981 gcaccatttg tcatttgacg aatgaatctg atgggcacac aacatcgttg tatggtattg 2041 gatttggtcc cttcatcatt acaaacaagc acttgtttag aagaaataat ggaacactgt 2101 tggtccaatc actacatggt gtattcaagg tcaagaacac cacgactttg caacaacacc 2161 tcattgatgg gagggacatg ataattattc gcatgcctaa ggatttccca ccatttcctc 2221 aaaagctgaa atttagagag ccacaaaggg aagagcgcat atgtcttgtg acaaccaact 2281 tccaaactaa gagcatgtct agcatggtgt cagacactag ttgcacattc ccttcatctg 2341 atggcatatt ctggaagcat tggattcaaa ccaaggatgg gcagtgtggc aatccattag 2401 tatcaactag agatgggttc attgttggta tacactcagc atcgaatttc accaacacaa 2461 acaattattt cacaagcgtg ccgaaaaact tcatggaatt gttgacaaat caggaggcgc 2521 agcagtgggt tagtggttgg cgattaaatg ctgactcagt attgtggggg ggccataaag 2581 ttttcatggt gtaagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa 2641 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 2701 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 2761 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 2821 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 2881 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 2941 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 3001 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc 3061 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 3121 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 3181 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag ctagagctcg 3241 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 3301 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 3361 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 3421 caagggggag gattgggaag acaatagcag gcatgctggg gaggtaacca cgtgcggacc 3481 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg 3541 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca 3601 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg 3661 catctgtgcg gtattcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg 3721 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 3781 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 3841 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 3901 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac 3961 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 4021 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat 4081 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa 4141 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata 4201 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct 4261 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt 4321 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata 4381 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt 4441 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 4501 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 4561 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 4621 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 4681 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 4741 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 4801 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 4861 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 4921 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 4981 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 5041 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 5101 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 5161 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 5221 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc 5281 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 5341 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 5401 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 5461 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 5521 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 5581 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 5641 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 5701 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa 5761 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 5821 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 5881 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 5941 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 6001 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 6061 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 6121 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 6181 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt //