LOCUS Exported 5954 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PPI reporter. ACCESSION . VERSION . KEYWORDS PCDNA3-GFP1-9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5954) AUTHORS Zhang Q, Zheng YW, Coughlin SR, Shu X TITLE A rapid fluorogenic GPCR-beta-arrestin interaction assay. JOURNAL Protein Sci. 2018 Apr;27(4):874-879. doi: 10.1002/pro.3385. Epub 2018 Feb 23. PUBMED 29411438 REFERENCE 2 (bases 1 to 5954) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5954 /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" primer_bind complement(1311..1328) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1317..1541 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1587..2015 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1674..1693) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1884..1905 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2024..2044) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2029..2358 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2209..2344 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2271..2290 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2425..3219 /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(2479..2498) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3089..3108 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3393..3514 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3430..3449) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3484..3503 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3563..3579) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3563..3579) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3576..3598) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3587..3603 /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(3611..3641) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3656..3677 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3794..3811) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3965..4553) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4045..4064) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4724..5584) /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 5347..5366 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5585..5689) /gene="bla" /label=AmpR promoter primer_bind complement(5764..5783) /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 ttgccaccat gcgcaaaggc gaagaactgt ttaccggcat 721 tgtgccgatt ctggtggaac tggatggcga tgtgaacggc cataaatttt ttgtgcgcgg 781 cgaaggcgaa ggcgatgcga ccattggcaa actgagcctg aaatttattt gcaccaccgg 841 caaactgccg gtgccgtggc cgaccctggt gaccaccctg acctatggcg tgcagtgctt 901 tagccgctat ccggatcata tgaaacgcca tgattttttt aaaagcgcga tgccggaagg 961 ctatgtgcag gaacgcacca tttattttaa agatgatggc acctataaaa cccgcgcgga 1021 agtgaaattt gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggca ttgattttaa 1081 agaagatggc aacattctgg gccataaact ggaatataac tttaacagcc ataaagtgta 1141 tattaccgcg gataaacaga acaacggcat taaagcgaac tttaccattc gccataacgt 1201 ggaagatggc agcgtgcagc tggcggatca ttatcagcag aacaccccga ttggcgatgg 1261 cccggttctt cttccttaag aattcgggcc cgtttaaacc cgctgatcag cctcgactgt 1321 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 1381 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 1441 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 1501 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 1561 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 1621 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 1681 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 1741 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 1801 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 1861 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 1921 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 1981 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 2041 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 2101 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 2161 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 2221 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 2281 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 2341 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 2401 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 2461 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 2521 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 2581 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 2641 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 2701 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 2761 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 2821 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 2881 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 2941 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 3001 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 3061 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 3121 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 3181 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 3241 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 3301 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 3361 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 3421 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 3481 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 3541 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 3601 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 3661 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 3721 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 3781 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 3841 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 3901 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 3961 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 4021 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 4081 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 4141 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 4201 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 4261 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 4321 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 4381 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 4441 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 4501 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 4561 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 4621 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 4681 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 4741 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 4801 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 4861 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 4921 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 4981 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 5041 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 5101 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 5161 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 5221 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 5281 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 5341 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 5401 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 5461 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 5521 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 5581 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 5641 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 5701 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct 5761 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt 5821 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga 5881 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc 5941 cagatatacg cgtt //