LOCUS Exported 8714 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of G protein-coupled receptors for PRESTO-Tango: parallel receptorome expression and screening via transcriptional output, with transcriptional activation following arrestin translocation. ACCESSION . VERSION . KEYWORDS GPR56-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8714) AUTHORS Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM, Sciaky N, Roth BL TITLE PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome. JOURNAL Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. Epub 2015 Apr 20. PUBMED 25895059 REFERENCE 2 (bases 1 to 8714) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8714 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 3038..4039 /codon_start=1 /product="tetracycline-controlled transactivator, comprising a fusion of the tetracycline repressor TetR with the C-terminal activation domain of herpes simplex virus VP16" /label=tTA /note="In the Tet-Off(R) system, tTA binds to the Tet-responsive element and stimulates transcription only in the absence of tetracycline or doxycycline." /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIEG LLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHADA LDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG" primer_bind complement(3105..3124) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(4071..4088) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4077..4301 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4347..4775 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4434..4453) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4644..4665 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4784..4804) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4789..5118 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4969..5104 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5031..5050 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5185..5979 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5239..5258) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5849..5868 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6153..6274 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6190..6209) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6244..6263 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6323..6339) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6323..6339) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6336..6358) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6347..6363 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6371..6401) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6416..6437 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6554..6571) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6725..7313) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6805..6824) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7484..8344) /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 8107..8126 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8345..8449) /gene="bla" /label=AmpR promoter primer_bind complement(8524..8543) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa 721 ttctgcagat atccagcaca gtggcggccg cgccaccatg aagacgatca tcgccctgag 781 ctacatcttc tgcctggtat tcgccgacta caaggacgat gatgacgcca gcatcgatat 841 gacaccacag agcctcctgc aaacgaccct tttcctcctg tcactgctct tcttggtcca 901 aggggctcat gggcggggac atcgcgagga tttcagattt tgcagtcagc gcaatcaaac 961 acaccgaagt agtctccact acaagcccac acctgacctg cggatctcca tcgaaaactc 1021 cgaggaggcc ctgaccgtcc atgctccttt tcctgccgcc catcctgcct ctcggagctt 1081 tcctgatccc cgcgggcttt accacttctg cctgtattgg aacaggcacg ccggccgctt 1141 gcacctgctc tacggaaaac gggattttct gctgtctgac aaagcctctt ctctgctttg 1201 tttccagcac caggaggaaa gtcttgccca gggaccaccc ctcctggcca cctcagttac 1261 ctcatggtgg agccctcaga acatctctct tcccagtgct gcctccttta cattcagttt 1321 tcacagcccc ccccatacag ccgcccacaa cgcctcagtg gacatgtgtg aactgaagcg 1381 ggacctccag ctgctttcac agtttcttaa gcatccccag aaggcctctc gacgacccag 1441 tgcggcaccg gctagtcagc agctccaatc cctggaatca aagctgacta gcgtaaggtt 1501 catgggtgac atggtgagtt tcgaggagga taggataaat gctaccgtgt ggaagctgca 1561 gccgaccgcc gggctgcagg accttcacat ccacagtcgc caggaagaag aacagagcga 1621 gattatggaa tacagtgtcc ttctcccgag gacgctgttt cagaggacga aaggcaggtc 1681 cggggaagcc gagaagcgcc tcctgctcgt ggacttctct agccaagccc tgtttcaaga 1741 caagaattct agtcaggttt tgggagaaaa agtactggga atcgtcgtgc agaataccaa 1801 ggtggccaat ctgaccgagc ccgtggtcct cacatttcag catcagctgc agcccaagaa 1861 cgtgaccctg caatgcgtgt tttgggtgga agaccctact ctttcaagcc ccggccattg 1921 gagttctgct ggctgcgaaa ctgtgcggag agagacacaa acttcctgtt tctgtaatca 1981 cctcacatat tttgccgtcc ttatggtttc atctgtagaa gtcgacgccg tccataaaca 2041 ctacctttcc ttgctgagct atgtggggtg tgttgtcagc gctctcgcct gtctggtgac 2101 aatcgcagcg tacctgtgca gccgggtgcc actgccgtgc cggcgcaaac ctagggacta 2161 taccataaag gtacacatga atttgctcct tgcagttttc ctcctggata cctcctttct 2221 gctcagtgag ccagtggcgt tgaccgggag cgaggcaggg tgccgagcga gtgctatctt 2281 cctccatttc tctcttctga cttgccttag ttggatggga ctggaaggct acaatcttta 2341 caggctggtg gtcgaggtgt ttggaaccta tgtacccggc tacctgctga agctgtccgc 2401 tatgggctgg ggttttccca tattcctcgt tacactggtc gccctggttg atgtagataa 2461 ttacggacca attattcttg ccgtgcatag gaccccggag ggcgtaatct atcccagcat 2521 gtgctggatc agggattcac ttgtttctta catcacaaat cttggactct ttagcttggt 2581 ctttcttttc aacatggcaa tgctggctac catggtggtc cagatcctgc gcctgagacc 2641 ccatacacag aagtggagcc acgtacttac tttgctcggc ctgtcacttg tcctcggcct 2701 tccttgggcc ctgatattct tctcattcgc tagcgggacc ttccagctgg tagtcctcta 2761 cttgttttcc attatcacat ctttccaggg ctttctcatt ttcatctggt attggagcat 2821 gagactgcag gcccggggtg gtcctagccc gttgaagtcc aatagcgact ctgctcgact 2881 tcctatctcc agcggttcca catcttccag tcgcatcatc gataccggtg gacgcacccc 2941 acccagcctg ggtccccaag atgagtcctg caccaccgcc agctcctccc tggccaagga 3001 cacttcatcg accggtgaga acctgtactt ccagctaaga ttagataaaa gtaaagtgat 3061 taacagcgca ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact 3121 cgcccagaag ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc 3181 tttgctcgac gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt 3241 agaaggggaa agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt 3301 actaagtcat cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta 3361 tgaaactctc gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc 3421 attatatgca ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga 3481 gcatcaagtc gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt 3541 acgacaagct atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct 3601 tgaattgatc atatgcggat tagaaaaaca acttaaatgt gaaagtgggt ccgcgtacag 3661 ccgcgcgcgt acgaaaaaca attacgggtc taccatcgag ggcctgctcg atctcccgga 3721 cgacgacgcc cccgaagagg cggggctggc ggctccgcgc ctgtcctttc tccccgcggg 3781 acacacgcgc agactgtcga cggccccccc gaccgatgtc agcctggggg acgagctcca 3841 cttagacggc gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt tcgatctgga 3901 catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact ccgcccccta 3961 cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg cccttggaat 4021 tgacgagtac ggtgggtagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt 4081 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 4141 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 4201 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 4261 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 4321 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 4381 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 4441 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 4501 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 4561 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 4621 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 4681 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 4741 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 4801 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 4861 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 4921 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 4981 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 5041 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 5101 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 5161 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 5221 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 5281 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 5341 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 5401 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 5461 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 5521 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 5581 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 5641 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 5701 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 5761 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 5821 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 5881 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 5941 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 6001 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 6061 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 6121 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 6181 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 6241 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 6301 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 6361 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 6421 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 6481 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 6541 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 6601 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 6661 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 6721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 6781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 6841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 6901 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 6961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 7141 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 7201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 7261 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7321 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7381 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 7441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 7501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 7561 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 7621 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 7681 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 7741 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 7801 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 7861 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 7921 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 7981 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8041 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8101 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 8161 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 8221 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8281 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8341 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 8401 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 8461 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct 8521 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt 8581 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga 8641 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc 8701 cagatatacg cgtt //