LOCUS Exported 6596 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 CCR4-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6596) 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 6596) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6596 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2190..2207) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2196..2420 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2466..2894 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2553..2572) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2763..2784 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2903..2923) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2908..3237 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3088..3223 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3150..3169 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3304..4098 /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(3358..3377) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3968..3987 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4272..4393 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4309..4328) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4363..4382 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4442..4458) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4442..4458) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4455..4477) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4466..4482 /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(4490..4520) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4535..4556 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4673..4690) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4844..5429) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4924..4943) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5600..6460) /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 6223..6242 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6461..6565) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc 961 agatatccag cacagtggcg gccgcgccac catgaagacg atcatcgccc tgagctacat 1021 cttctgcctg gtattcgccg actacaagga cgatgatgac gccagcatcg atatgaatcc 1081 cactgacatc gcagatacca ctttggacga gagtatctac agcaattact acttgtacga 1141 aagcatccca aagccgtgca ctaaggaggg cattaaagcg ttcggggagc ttttcctccc 1201 ccccttgtac tccctggtgt ttgtcttcgg cttgttgggt aactccgttg tggtattggt 1261 attgttcaag tacaagagac ttaggagcat gaccgacgtt tacttgctga atctggcaat 1321 tagcgatctt cttttcgtat tttccctgcc cttttggggg tattatgcgg ccgaccagtg 1381 ggtgttcgga ctcggcctgt gtaagatgat ttcttggatg taccttgtgg ggttctactc 1441 agggatcttt ttcgtgatgc tcatgtctat tgatagatat ttggccattg tgcatgctgt 1501 cttttctctt cgggcaagga ctctcaccta tggggttatt acctccctgg ccacgtggtc 1561 agtggcagtt ttcgcctcac tccccggatt cctcttcagc acgtgctaca cagagcgcaa 1621 ccatacgtat tgtaagacca agtacagcct caatagcaca acatggaagg tgctttctag 1681 tctggagatt aatatcctgg ggcttgtgat tccgctgggc atcatgctct tttgctacag 1741 catgataatt agaacactgc agcattgtaa aaacgagaaa aaaaacaagg cggtcaagat 1801 gatcttcgca gtggtagtac tgttcctggg gttctggaca ccttataaca tagtactctt 1861 tttggaaaca ctggtagaac tcgaagtact gcaggattgc accttcgagc gctacctgga 1921 ttacgccatc caggctaccg agacacttgc cttcgtacac tgttgtctta atccaattat 1981 ctactttttc cttggcgaaa agttcagaaa gtacattttg cagctcttta agacctgccg 2041 gggtctcttt gtgttgtgtc aatattgcgg gcttctccag atctacagcg cggacacacc 2101 ctcaagctct tatacacaga gtacaatgga ccatgacctg catgacgccc tcatcgagtc 2161 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 2221 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 2281 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2341 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 2401 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 2461 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2521 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2581 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2641 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2701 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2761 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 2821 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 2881 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3061 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3121 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3181 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3241 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3301 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3361 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3421 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3481 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3541 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3601 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3661 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3721 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3781 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 3841 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 3901 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 3961 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4021 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4081 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4141 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4201 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4261 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4321 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 4381 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 4441 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4501 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4681 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 5041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 5161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 5221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 5281 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 5341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc 5401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 5821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 6241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 6301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //