LOCUS Exported 6012 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION StrepIII-HA-tagged gene expression. ACCESSION . VERSION . KEYWORDS GRB2-SHN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6012) AUTHORS Yadav L, Pietila E, Ohman T, Liu X, Mahato AK, Sidorova Y, Lehti K, Saarma M, Varjosalo M TITLE PTPRA Phosphatase Regulates GDNF-Dependent RET Signaling and Inhibits the RET Mutant MEN2A Oncogenic Potential. JOURNAL iScience. 2020 Feb 21;23(2):100871. doi: 10.1016/j.isci.2020.100871. Epub 2020 Jan 31. PUBMED 32062451 REFERENCE 2 (bases 1 to 6012) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6012 /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" protein_bind 820..838 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 841..859 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 869..893 /label=LNCX /note="Human CMV promoter, forward primer" CDS 1064..1090 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1121..1144 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 1181..1204 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" protein_bind 1217..1241 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(1902..1926) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(1964..1981) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1970..2194 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2327..2346) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2478..2525 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 2533..3555 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 3685..3806 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3722..3741) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3776..3795 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3855..3871) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3855..3871) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3868..3890) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3879..3895 /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(3903..3933) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3948..3969 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4086..4103) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4257..4845) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4337..4356) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5016..5876) /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 5639..5658 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5877..5981) /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 ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga 1021 attctgcaga tatccagcac agtggcggcc gctcgagacc atgtacccat acgatgttcc 1081 tgactatgcc ggtaccgagc tcggatccac catggctagc tggagccacc cgcagttcga 1141 gaaaggtgga ggttccggag gtggatcggg aggtggatcg tggagccacc cgcagttcga 1201 aaaagcggcc gatatcacaa gtttgtacaa aaaagcaggc tccaccatgg aagccatcgc 1261 caaatatgac ttcaaagcta ctgcagacga cgagctgagc ttcaaaaggg gggacatcct 1321 caaggttttg aacgaagaat gtgatcagaa ctggtacaag gcagagctta atggaaaaga 1381 cggcttcatt cccaagaact acatagaaat gaaaccacat ccgtggtttt ttggcaaaat 1441 ccccagagcc aaggcagaag aaatgcttag caaacagcgg cacgatgggg cctttcttat 1501 ccgagagagt gagagcgctc ctggggactt ctccctctct gtcaagtttg gaaacgatgt 1561 gcagcacttc aaggtgctcc gagatggagc cgggaagtac ttcctctggg tggtgaagtt 1621 caattctttg aatgagctgg tggattatca cagatctaca tctgtctcca gaaaccagca 1681 gatattcctg cgggacatag aacaggtgcc acagcagccg acatacgtcc aggccctctt 1741 tgactttgat ccccaggagg atggagagct gggcttccgc cggggagatt ttatccatgt 1801 catggataac tcagacccca actggtggaa aggagcttgc cacgggcaga ccggcatgtt 1861 tccccgcaat tatgtcaccc ccgtgaaccg gaacgtcttg gacccagctt tcttgtacaa 1921 agtggtgacg taagctaggg gcccgtttaa acccgctgat cagcctcgac tgtgccttct 1981 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc 2041 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt 2101 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat 2161 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg 2221 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 2281 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 2341 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 2401 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 2461 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga 2521 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa 2581 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc 2641 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc 2701 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg 2761 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt 2821 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag 2881 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga 2941 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc 3001 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct 3061 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg 3121 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg 3181 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg 3241 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca 3301 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc 3361 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc 3421 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg 3481 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg 3541 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg 3601 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 3661 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 3721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 3781 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 3841 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 3901 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 3961 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 4021 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 4081 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 4141 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 4201 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 4261 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 4321 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 4381 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 4441 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 4501 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 4561 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 4621 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 4681 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 4741 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 4801 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 4861 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 4921 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 4981 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 5041 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 5101 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 5161 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 5221 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 5281 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 5341 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 5401 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 5461 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 5521 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 5581 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 5641 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 5701 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 5761 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 5821 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 5881 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 5941 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 6001 ccacctgacg tc //