LOCUS Exported 6414 bp ds-DNA circular SYN 17-JAN-2024 DEFINITION Mammalian expression of HA tagged human p62. ACCESSION . VERSION . KEYWORDS HA-p62 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6414) AUTHORS Fan W, Tang Z, Chen D, Moughon D, Ding X, Chen S, Zhu M, Zhong Q TITLE Keap1 facilitates p62-mediated ubiquitin aggregate clearance via autophagy. JOURNAL Autophagy. 2010 Jul 22. 6(5):. PUBMED 20495340 REFERENCE 2 (bases 1 to 6414) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 17, 2024 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6414 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(13..33) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 18..347 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 198..333 /label=SV40 ori /note="SV40 origin of replication" primer_bind 260..279 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 395..442 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 461..835 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 965..1086 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1002..1021) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1056..1075 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1135..1151) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1135..1151) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1148..1170) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1159..1175 /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(1183..1213) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1228..1249 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1366..1383) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1537..2125) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1617..1636) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2296..3156) /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 2919..2938 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3157..3261) /gene="bla" /label=AmpR promoter primer_bind complement(3336..3355) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 3527..3906 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3907..4110 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4061..4081 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 4112..4130 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4133..4151 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 4161..4185 /label=LNCX /note="Human CMV promoter, forward primer" CDS 5647..5673 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(5714..5731) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5720..5944 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5990..4 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6077..6096) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6287..6308 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 61 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 361 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta 421 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc 481 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct 541 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac 601 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg 661 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg 721 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc 781 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt 841 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 1021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 1081 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 1141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 1201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 1261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 1321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 1381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 1441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 1501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 1561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 1621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 1681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 1741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 1801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 1861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 1921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 1981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 2041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 2101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 2161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 2221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 2281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 2341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 2401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 2461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 2521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 2581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 2641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 2701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 2761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 2821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 2881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 2941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 3001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 3061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 3121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 3181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 3241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc 3301 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag 3361 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa 3421 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt 3481 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg 3541 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc 3601 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 3661 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 3721 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 3781 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 3841 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 3901 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg 3961 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa 4021 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt 4081 gtacggtggg aggtctatat aagcagagct ctccctatca gtgatagaga tctccctatc 4141 agtgatagag atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg gagacgccat 4201 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcg 4261 tttaaactta agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattctatg 4321 gcgtcgctca ccgtgaaggc ctaccttctg ggcaaggagg acgcggcgcg cgagattcgc 4381 cgcttcagct tctgctgcag ccccgagcct gaggcggaag ccgaggctgc ggcgggtccg 4441 ggaccctgcg agcggctgct gagccgggtg gccgccctgt tccccgcgct gcggcctggc 4501 ggcttccagg cgcactaccg cgatgaggac ggggacttgg ttgccttttc cagtgacgag 4561 gaattgacaa tggccatgtc ctacgtgaag gatgacatct tccgaatcta cattaaagag 4621 aaaaaagagt gccggcggga ccaccgccca ccgtgtgctc aggaggcgcc ccgcaacatg 4681 gtgcacccca atgtgatctg cgatggctgc aatgggcctg tggtaggaac ccgctacaag 4741 tgcagcgtct gcccagacta cgacttgtgt agcgtctgcg agggaaaggg cttgcaccgg 4801 gggcacacca agctcgcatt ccccagcccc ttcgggcacc tgtctgaggg cttctcgcac 4861 agccgctggc tccggaaggt gaaacacgga cacttcgggt ggccaggatg ggaaatgggt 4921 ccaccaggaa actggagccc acgtcctcct cgtgcagggg aggcccgccc tggccccacg 4981 gcagaatcag cttctggtcc atcggaggat ccgagtgtga atttcctgaa gaacgttggg 5041 gagagtgtgg cagctgccct tagccctctg ggcattgaag ttgatatcga tgtggagcac 5101 ggagggaaaa gaagccgcct gacccccgtc tctccagaga gttccagcac agaggagaag 5161 agcagctcac agccaagcag ctgctgctct gatcccagca agccgggtgg gaatgttgag 5221 ggcgccacgc agtctctggc ggagcagatg agaaagatcg ccttggagtc cgaggggcgc 5281 cctgaggaac agatggagtc ggataactgt tcaggaggag atgatgactg gacccatctg 5341 tcttcaaaag aagtggaccc gtctacaggt gaactccagt ccctacagat gccagaatcc 5401 gaagggccaa gctctctgga cccctcccag gagggaccca cagggctgaa ggaagctgcc 5461 ttgtacccac atctcccgcc agaggctgac ccgcggctga ttgagtccct ctcccagatg 5521 ctgtccatgg gcttctctga tgaaggcggc tggctcacca ggctcctgca gaccaagaac 5581 tatgacatcg gagcggctct ggacaccatc cagtattcaa agcatccccc gccgttgcgc 5641 ggccgctacc catacgatgt tccagattac gcttgactcg agtctagagg gcccgtttaa 5701 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc 5761 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 5821 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag 5881 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct 5941 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt 6001 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 6061 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 6121 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 6181 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 6241 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 6301 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 6361 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcg //