LOCUS Exported 6274 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses SPARK2 NanoLuc-beta-arrestin2-TEVp (no HA) in mammalian cells. ACCESSION . VERSION . KEYWORDS pAAV-NanoLuc-15aa linker-beta-arrestin2-no HA-GS linker-TEVp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6274) TITLE Luciferase-LOV BRET enables versatile and specific transcriptional readout of cellular protein-protein interactions JOURNAL Unpublished REFERENCE 2 (bases 1 to 6274) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6274 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" enhancer 201..504 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 505..708 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 659..679 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 753..772 /label=T7 /note="T7 promoter, forward primer" promoter 753..771 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 785..794 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 791..1303 /codon_start=1 /product="NanoLuc(R) luciferase" /label=Nluc /note="human codon-optimized" /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW RLCERILA" primer_bind complement(3296..3313) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3302..3509 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3538..3678 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3538..3667 /label=AAV2 ITR rep_origin 3752..4207 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3839..3858) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4049..4070 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4224..4243) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4343..4365 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4403..4421) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4489..4593 /gene="bla" /label=AmpR promoter CDS 4594..5454 /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(4812..4831) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5625..6213 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6114..6133 /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 tagtaatcaa ttacggggtc attagttcat 181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg 241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 301 gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta 361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact 721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa 781 gcttgccacc atggtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg 841 ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg 901 ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga 961 catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa 1021 aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg 1081 cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga 1141 aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa 1201 caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat 1261 caacggagtg accggctggc ggctgtgcga acgcattctg gcgggcgacc cactagtcca 1321 gtgtggtgga attgccggat ccgccaccgg tgaaaaaccc gggaccaggg tcttcaagaa 1381 gtcgagccct aactgcaagc tcaccgtgta cttgggcaag cgtgactttg tggatcactt 1441 ggacaaagtg gatcctgtcg atggtgtggt gcttgtggat cctgactact tgaaggaccg 1501 gaaagtgttt gtgaccctca cctgtgcctt ccgctatggc cgagaagacc tggatgtact 1561 gggcctgtct ttccgcaaag atctgttcat cgccacctac caggccttcc cccccatgcc 1621 caacccacct cggcccccca cccgcctaca ggaccgactg ctgaagaagt tgggccagca 1681 tgcccacccc ttttttttca caatacccca gaatttgcct tgctccgtca cactgcagcc 1741 aggaccggag gacacaggga aggcctgtgg agtagacttt gagattcgag ccttctgtgc 1801 caaatctata gaagaaaaaa gccacaaaag gaactccgtg cggcttatca tcagaaaggt 1861 acagtttgct cctgagacac ccggccccca gccatcagct gaaaccacac gccacttcct 1921 catgtctgac cggaggtccc tgcacctaga ggcttccctg gacaaagagc tgtactacca 1981 tggggaaccc ctcaatgtca acgtccacgt caccaacaat tctgccaaga ccgtcaagaa 2041 gatcagagtg tctgtgagac agtatgccga catttgcctc ttcagcaccg cgcagtacaa 2101 gtgtcctgtg gctcagcttg aacaagatga ccaggtgtct cccagttcca cattctgcaa 2161 ggtgtacacc ataaccccgc tgctcagtga caaccgagag aagcgtggcc ttgcccttga 2221 tgggcaactc aagcacgaag acaccaacct ggcttccagc accattgtga aggagggagc 2281 caacaaggag gtgctgggaa tcctagtatc ctacagggtc aaggtgaagc tggtggtgtc 2341 tcgaggcggg gatgtctccg tggagctacc tttcgtccta atgcacccca agccccacga 2401 ccacatcacc cttccccgac cccagtcagc cccccgggaa atagacatcc ctgtggatac 2461 caacctcatt gaattcgata ccaactatgc cacagacgac gacatcgtgt ttgaggactt 2521 tgcgaggctt cggctgaagg ggatgaagga tgacgactgt gatgaccagt tctgcggtgg 2581 aagtggatca ggcagcggtg gatctggaga aagcttgttt aagggaccac gtgattacaa 2641 cccgatatcg agcaccattt gtcatttgac gaatgaatct gatgggcaca caacatcgtt 2701 gtatggtatt ggatttggtc ccttcatcat tacaaacaag cacttgttta gaagaaataa 2761 tggaacactg ttggtccaat cactacatgg tgtattcaag gtcaagaaca ccacgacttt 2821 gcaacaacac ctcattgatg ggagggacat gataattatt cgcatgccta aggatttccc 2881 accatttcct caaaagctga aatttagaga gccacaaagg gaagagcgca tatgtcttgt 2941 gacaaccaac ttccaaacta agagcatgtc tagcatggtg tcagacacta gttgcacatt 3001 cccttcatct gatggcatat tctggaagca ttggattcaa accaaggatg ggcagtgtgg 3061 cagtccatta gtatcaacta gagatgggtt cattgttggt atacactcag catcgaattt 3121 caccaacaca aacaattatt tcacaagcgt gccgaaaaac ttcatggaat tgttgacaaa 3181 tcaggaggcg cagcagtggg ttagtggttg gcgattaaat gctgactcag tattgtgggg 3241 tggccataaa gttttcatgg tgtaagaatt cctcgagcta gagctcgctg atcagcctcg 3301 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 3361 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 3421 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 3481 tgggaagaca atagcaggca tgctggggag gtaaccacgt gcggaccgag cggccgcagg 3541 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg 3601 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag 3661 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 3721 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg 3781 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 3841 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 3901 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 3961 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 4021 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 4081 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg 4141 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt 4201 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc 4261 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 4321 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 4381 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 4441 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 4501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 4561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 4621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 4681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 4741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact 4801 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc 4861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 4921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 4981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 5041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 5101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc 5161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 5221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 5281 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca 5341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 5401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 5461 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 5521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 5581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 5641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 5701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 5761 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 5821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 5881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 5941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 6001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 6061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 6121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 6181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 6241 ttcctggcct tttgctggcc ttttgctcac atgt //