LOCUS Exported 7609 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses SEC62 in mammalian cells. ACCESSION . VERSION . KEYWORDS pMRX-INU-SEC62-FLAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7609) AUTHORS Chino H, Hatta T, Natsume T, Mizushima N TITLE Intrinsically Disordered Protein TEX264 Mediates ER-phagy. JOURNAL Mol Cell. 2019 Apr 11. pii: S1097-2765(19)30257-6. doi: 10.1016/j.molcel.2019.03.033. PUBMED 31006538 REFERENCE 2 (bases 1 to 7609) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7609 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" misc_feature 739..1096 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1161..1577 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1507..1529 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1545..1561 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 1587..1961 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind 1899..1915 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" CDS 3226..3291 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" misc_feature 3341..3893 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3485..3502) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3712..3731 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 3911..4705 /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(3965..3984) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4575..4594 /label=Neo-F /note="Neomycin resistance gene, forward primer" LTR 4935..5404 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(5534..5551) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5705..6293) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5785..5804) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6457..7317) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7080..7099 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7318..7422) /gene="bla" /label=AmpR promoter primer_bind 7490..7508 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" 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 ctagagaacc atcagatgtt tccagggtgc cccaaggacc 421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc 481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc 541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat 601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca 661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc 721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt 781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg 841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc 901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg 961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc 1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg 1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt 1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc 1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt 1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca 1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg 1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc 1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt 1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca 1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc 1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc 1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc 1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg 1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc 1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg 1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatat 1981 aaagcttaaa ggatccggga attcatggcg gaacgcagga gacacaagaa gcggatccag 2041 gaagttggtg aaccatctaa agaagagaag gctgtggcca agtatcttcg attcaactgt 2101 ccaacaaagt ccaccaatat gatgggtcac cgggttgatt attttattgc ttcaaaagca 2161 gtggactgtc ttttggattc aaagtgggca aaggccaaga aaggagagga agctttattt 2221 acaaccaggg agtctgtggt tgactactgc aacaggcttt taaagaagca gttttttcac 2281 cgagccctaa aagtaatgaa aatgaaatat gataaagaca taaagaaaga aaaagataaa 2341 ggaaaagctg aaagtggaaa agaagaagat aaaaagagca agaaagaaaa tataaaggat 2401 gagaagacaa aaaaagaaaa agagaaaaaa aaagatggtg aaaaggaaga atccaaaaag 2461 gaggaaactc caggaactcc taaaaagaag gaaactaaga aaaaattcaa acttgagcca 2521 catgatgatc aggtttttct ggatggaaat gaggtgtatg tatggatcta tgacccagtt 2581 cactttaaaa catttgtcat gggattaatt cttgtgattg cagtaatagc ggccaccctc 2641 ttcccccttt ggccagcaga aatgagagta ggtgtttatt acctcagtgt gggtgcaggc 2701 tgttttgtag ccagtattct tctccttgct gttgctcgat gcattctatt tctcatcatt 2761 tggctcataa ctggaggaag gcaccacttt tggttcttgc caaatctgac tgctgatgtg 2821 ggcttcattg actccttcag gcctctgtac acacatgaat acaaaggacc aaaagcagac 2881 ttaaagaaag atgagaagtc tgaaaccaaa aagcaacaga agtccgacag tgaggaaaag 2941 tcagacagtg agaaaaagga agatgaggag gggaaagtag gaccaggaaa tcatggaaca 3001 gaaggctcgg ggggagaacg gcattcagac acggacagtg acaggaggga agatgatcga 3061 tcccagcaca gtagtggaaa tggaaatgat tttgaaatga taacaaaaga ggaactggaa 3121 cagcaaacag atggggattg tgaagaggat gaggaagagg aaaatgatgg agaaacacct 3181 aaatcttcac atgaaaaatc agcgaattcg ggctctggtt caggcgacta caaagaccat 3241 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtaatgattt 3301 ctcgagagcg gccgctacgt aaattccgcc ccccccccta acgttactgg ccgaagccgc 3361 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt 3421 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt 3481 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg 3541 gaagtttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca 3601 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg 3661 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc 3721 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct 3781 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta 3841 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac 3901 aaccgtcgac atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga 3961 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt 4021 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct 4081 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg 4141 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt 4201 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc 4261 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc 4321 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga 4381 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg 4441 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat 4501 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg 4561 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc 4621 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta 4681 tcgccttctt gacgagttct tctgagtcga ccgcccgccc cacgacccgc agcgcccgac 4741 cgaaaggagc gcacgacccc atggctcgac gataaaataa aagattttat ttagtctcca 4801 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc 4861 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg 4921 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc 4981 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt 5041 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc 5101 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc 5161 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc 5221 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga 5281 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg 5341 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct 5401 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat 5461 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5521 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5581 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5641 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5701 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5761 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5821 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5881 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5941 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 6001 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 6061 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 6121 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 6181 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 6241 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 6301 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 6361 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg gccgcaaatc 6421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6541 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6601 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 6661 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6721 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6781 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6841 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6901 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6961 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 7021 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga 7081 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 7141 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7201 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 7261 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 7321 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 7381 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7441 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat 7501 cacgaggccc tttcgtcttc actataggag cttggcccat tgcatacgtt gtatccatat 7561 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgtt //