LOCUS Exported 6329 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses SPARK2 NanoLuc-beta-arrestin2-TEVp in mammalian cells. ACCESSION . VERSION . KEYWORDS pAAV-NanoLuc-15aa linker-beta-arrestin2-HA-GS linker-TEVp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6329) TITLE Luciferase-LOV BRET enables versatile and specific transcriptional readout of cellular protein-protein interactions JOURNAL Unpublished REFERENCE 2 (bases 1 to 6329) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6329 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 6..135 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" enhancer 206..509 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 510..713 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 664..684 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 758..777 /label=T7 /note="T7 promoter, forward primer" promoter 758..776 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 790..799 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 796..1308 /codon_start=1 /product="NanoLuc(R) luciferase" /label=Nluc /note="human codon-optimized" /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW RLCERILA" CDS 2581..2607 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2608..2634 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(3355..3372) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3361..3568 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3597..3737 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3597..3726 /label=AAV2 ITR rep_origin 3812..4267 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3899..3918) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4109..4130 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4284..4303) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4403..4425 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4463..4481) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4549..4653 /gene="bla" /label=AmpR promoter CDS 4654..5514 /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(4872..4891) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5685..6273 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6174..6193 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga 61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca 121 tcactagggg ttcctgcggc cgcacgcgtt gtacatagta atcaattacg gggtcattag 181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct 241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc 301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg 361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat 421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc 721 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga 781 cccaagcttg ccaccatggt cttcacactc gaagatttcg ttggggactg gcgacagaca 841 gccggctaca acctggacca agtccttgaa cagggaggtg tgtccagttt gtttcagaat 901 ctcggggtgt ccgtaactcc gatccaaagg attgtcctga gcggtgaaaa tgggctgaag 961 atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc 1021 gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc actttaaggt gatcctgcac 1081 tatggcacac tggtaatcga cggggttacg ccgaacatga tcgactattt cggacggccg 1141 tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg taacagggac cctgtggaac 1201 ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg gctccctgct gttccgagta 1261 accatcaacg gagtgaccgg ctggcggctg tgcgaacgca ttctggcggg cgacccacta 1321 gtccagtgtg gtggaattgc cggatccgcc accggtgaaa aacccgggac cagggtcttc 1381 aagaagtcga gccctaactg caagctcacc gtgtacttgg gcaagcgtga ctttgtggat 1441 cacttggaca aagtggatcc tgtcgatggt gtggtgcttg tggatcctga ctacttgaag 1501 gaccggaaag tgtttgtgac cctcacctgt gccttccgct atggccgaga agacctggat 1561 gtactgggcc tgtctttccg caaagatctg ttcatcgcca cctaccaggc cttccccccc 1621 atgcccaacc cacctcggcc ccccacccgc ctacaggacc gactgctgaa gaagttgggc 1681 cagcacgccc accccttttt tttcacaata ccccagaatt tgccttgctc cgtcacactg 1741 cagccaggac cggaggacac agggaaggcc tgtggagtag actttgagat tcgagccttc 1801 tgtgccaaat ctatagaaga aaaaagccac aaaaggaact ccgtgcggct tatcatcaga 1861 aaggtacagt ttgctcctga gacacccggc ccccagccat cagctgaaac cacacgccac 1921 ttcctcatgt ctgaccggag gtccctgcac ctagaggctt ccctggacaa agagctgtac 1981 taccatgggg aacccctcaa tgtcaacgtc cacgtcacca acaattctgc caaaaccgtc 2041 aagaagatca gagtgtctgt gagacagtat gccgacattt gcctcttcag caccgcgcag 2101 tacaagtgtc ctgtggctca gcttgaacaa gatgaccagg tgtctcccag ttccacattc 2161 tgcaaggtgt acaccataac cccgctgctc agtgacaacc gagagaagcg tggccttgcc 2221 cttgatgggc aactcaagca cgaagacacc aacctggctt ccagcaccat tgtgaaggag 2281 ggagccaaca aggaggtgct gggaatccta gtatcctaca gggtcaaggt gaagctggtg 2341 gtgtctcgag gcggggatgt ctccgtggag ctacctttcg tcctaatgca ccccaagccc 2401 cacgaccaca tcacccttcc ccgaccccag tcagcccccc gggaaataga catccctgtg 2461 gataccaacc tcattgaatt cgataccaac tatgccacag acgacgacat cgtgtttgag 2521 gactttgcga ggcttcggct gaaggggatg aaggatgacg actgtgatga ccagttctgc 2581 tacccatacg atgtgccaga ttacgcctat ccctatgacg tccctgatta cgctggtgga 2641 agtggatcag gcagcggtgg atctggagaa agcttgttta agggaccacg tgattacaac 2701 ccgatatcga gcaccatttg tcatttgacg aatgaatctg atgggcacac aacatcgttg 2761 tatggtattg gatttggtcc cttcatcatt acaaacaagc acttgtttag aagaaataat 2821 ggaacactgt tggtccaatc actacatggt gtattcaagg tcaagaacac cacgactttg 2881 caacaacacc tcattgatgg gagggacatg ataattattc gcatgcctaa ggatttccca 2941 ccatttcctc aaaagctgaa atttagagag ccacaaaggg aagagcgcat atgtcttgtg 3001 acaaccaact tccaaactaa gagcatgtct agcatggtgt cagacactag ttgcacattc 3061 ccttcatctg atggcatatt ctggaagcat tggattcaaa ccaaggatgg gcagtgtggc 3121 agtccattag tatcaactag agatgggttc attgttggta tacactcagc atcgaatttc 3181 accaacacaa acaattattt cacaagcgtg ccgaaaaact tcatggaatt gttgacaaat 3241 caggaggcgc agcagtgggt tagtggttgg cgattaaatg ctgactcagt attgtggggt 3301 ggccataaag ttttcatggt gtaagaattc ctcgagctag agctcgctga tcagcctcga 3361 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 3421 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 3481 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 3541 gggaagacaa tagcaggcat gctggggagg taaccacgtg cggaccgagc ggccgcagga 3601 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg 3661 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc 3721 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 3781 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg 3841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct 3901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 3961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 4021 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 4081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 4141 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg 4201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt 4261 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc 4321 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 4381 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 4441 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 4501 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 4561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 4621 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 4681 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 4741 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 4801 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact 4861 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc 4921 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 4981 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 5041 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 5101 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 5161 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc 5221 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 5281 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 5341 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca 5401 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 5461 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 5521 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 5581 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 5641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 5701 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 5761 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 5821 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 5881 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 5941 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 6001 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 6061 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 6121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 6181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 6241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 6301 ttcctggcct tttgctggcc ttttgctca //