LOCUS Exported 6272 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION b2AR-SPARK TF component. ACCESSION . VERSION . KEYWORDS pAAV-AVPR2-eLOV-TEVcs-FLAG-GAL4-V5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6272) AUTHORS Kim MW, Wang W, Sanchez MI, Coukos R, von Zastrow M, Ting AY TITLE Time-gated detection of protein-protein interactions with transcriptional readout. JOURNAL Elife. 2017 Nov 30;6. doi: 10.7554/eLife.30233. PUBMED 29189201 REFERENCE 2 (bases 1 to 6272) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6272 /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" CDS 2414..2437 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 2438..2875 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="DNA binding domain of the GAL4 transcriptional activator" /label=GAL4 DNA binding domain /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS" CDS 2876..3217 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="activation domain of the GAL4 transcriptional activator" /label=GAL4 activation domain /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD EDTPPNPKKE" CDS 3218..3259 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3293..3310) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3299..3506 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3535..3675 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3535..3664 /label=AAV2 ITR rep_origin 3750..4205 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3837..3856) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4047..4068 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4222..4241) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4341..4363 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4401..4419) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4487..4591 /gene="bla" /label=AmpR promoter CDS 4592..5452 /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(4810..4829) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5623..6211 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6112..6131 /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 atgggtggag 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 atgctcatgg cctctaccac aagcgctgtg cccggacatc catctcttcc 841 aagtcttccc tccaacagct cacaggagag gcctctggat acccgggacc cacttcttgc 901 aagagcggag ctcgccttgc tctccatcgt tttcgttgcc gtcgcgttga gcaatggtct 961 ggtgctggcc gccctggcgc gacgaggccg ccgaggccac tgggcgccta ttcatgtctt 1021 tatcggccat ctctgtcttg cagacctggc cgtggcgctg tttcaggtgc tccctcaact 1081 ggcttggaaa gccaccgata gattccgggg gccagacgca ctgtgtcggg ccgtgaaata 1141 cctgcagatg gtgggaatgt atgccagttc atatatgata cttgctatga ccttggacag 1201 acacagggct atctgtcgcc ccatgctggc gtataggcac ggttccggag cccattggaa 1261 caggccagtt ctggttgcgt gggctttttc cctgcttctg agcttgcccc agttgtttat 1321 tttcgcacag agaaatgtgg agggcggatc tggcgtgacc gattgctggg cttgcttcgc 1381 tgagccatgg ggcagaagga cctacgtgac gtggattgct ctcatggtgt tcgttgctcc 1441 caccctgggg atcgccgcat gccaggtcct gatattcaga gaaatacacg cctccttggt 1501 tcctgggcca agcgagcgcc ctggtggaag aagacggggt cgaaggactg gctctccagg 1561 cgaaggcgcc cacgtatcag ctgcggtagc taagaccgta cgcatgacac tggtcatcgt 1621 cgtcgtttac gttctttgct gggccccgtt ttttttggtg cagctctggg cggcctggga 1681 ccccgaagca cccctggaag gtgctccatt cgttctgctc atgcttctcg cttcactgaa 1741 tagctgtacc aatccttgga tttacgcctc tttctcctcc agtgttagct ccgagctgcg 1801 gagtcttctc tgttgcgccc ggggcaggac ccccccaagc ctgggtccac aggacgaatc 1861 ctgcactacc gcctccagtt ccctggccaa agacacctct tctgagctgg ccgagaagct 1921 ggccggcctg gacatcaacg gtggagctag cggcagccgg gcgaccaccc tggaacgtat 1981 tgaaaagagc tttgtgatta ccgatccgcg tctgccggat aacccgatta tttttgtgag 2041 cgatagcttt ctgcagctga ccgaatatag ccgtgaagaa attctgggcc gtaactgccg 2101 ttttctgcag ggcccggaaa ccgatcgtgc gaccgtgcgt aaaattcgtg atgcgattga 2161 taaccagacc gaagtgaccg tgcagctgat taactatacc aaaagcggca aaaaattttg 2221 gaacctgttt catctgcagc cgatgcgtga tcagaaaggc gatgtgcagt attttattgg 2281 cgtgcagctg gatggtaccg aacgtgtgcg tgatgcggcg gaacgtgaag ctgtgatgct 2341 ggtcaagaaa accgcggagg aaatcgacga agccgctaag gaaaatctct acttccagat 2401 gggcggagga tccgactaca aggatgacga cgataagaag ctactgtctt ctatcgaaca 2461 agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc 2521 caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct 2581 gactagggca catctgacag aagtggaatc aaggctagaa agactggaac agctatttct 2641 actgattttt cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat 2701 aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga 2761 tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc 2821 gacatcatca tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tatcggccaa 2881 ttttaatcaa agtgggaata ttgctgatag ctcattgtcc ttcactttca ctaacagtag 2941 caacggtccg aacctcataa caactcaaac aaattctcaa gcgctttcac aaccaattgc 3001 ctcctctaac gttcatgata acttcatgaa taatgaaatc acggctagta aaattgatga 3061 tggtaataat tcaaaaccac tgtcacctgg ttggacggac caaactgcgt ataacgcgtt 3121 tggaatcact acagggatgt ttaataccac tacaatggat gatgtatata actatctatt 3181 cgatgatgaa gataccccac caaacccaaa aaaagagggc aagccgatcc ctaaccctct 3241 gctgggcctg gacagcactt aagaattcct cgagctagag ctcgctgatc agcctcgact 3301 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 3361 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 3421 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 3481 gaagacaata gcaggcatgc tggggaggta accacgtgcg gaccgagcgg ccgcaggaac 3541 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc 3601 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc 3661 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt 3721 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc 3781 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc 3841 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 3901 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 3961 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 4021 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 4081 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt 4141 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac 4201 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg 4261 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta 4321 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc 4381 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat 4441 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat 4501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata 4561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct 4621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa 4681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa 4741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt 4801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg 4861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca 4921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa 4981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt 5041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc 5101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa 5161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga 5221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc 5281 tgataaatct ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga 5341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga 5401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga 5461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 5521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 5581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 5641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 5701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 5761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 5821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 5881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 5941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 6001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 6061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 6121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 6181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 6241 cctggccttt tgctggcctt ttgctcacat gt //