LOCUS Exported 9490 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses RTN3L in mammalian cells. ACCESSION . VERSION . KEYWORDS pMRX-INU-FLAG-RTN3L SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9490) 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 9490) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9490 /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" regulatory 1988..1997 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1997..2062 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS complement(2551..2562) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" misc_feature 5222..5774 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(5366..5383) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5593..5612 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5792..6586 /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(5846..5865) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6456..6475 /label=Neo-F /note="Neomycin resistance gene, forward primer" LTR 6816..7285 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(7415..7432) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7586..8174) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7666..7685) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8338..9198) /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 8961..8980 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9199..9303) /gene="bla" /label=AmpR promoter primer_bind 9371..9389 /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 aaagcttgcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga 2041 ctacaaggat gacgatgaca agggttcagg ctcgggatcc gggaattcga tggcggagcc 2101 gtcggcggcc actcagtccc attccatctc ctcgtcgtcc ttcggagccg agccgtccgc 2161 gcccggcggc ggcgggagcc caggagcctg ccccgccctg gggacgaaga gctgcagctc 2221 ctcctgtgcg gattcctttg tttcttcctc ttcctctcag cctgtatctc tattttcgac 2281 ctcacaagag ggattgagct ctctttgctc tgatgagcca tcttcagaaa ttatgacttc 2341 ttcctttctt tcatcttctg aaatacataa cactggcctt acaatactac atggagaaaa 2401 aagccatgtg ttagggagcc agcctatttt agccaaagaa ggaaaagacc acttggatct 2461 tctagatatg aaaaagatgg aaaagcctca ggggaccagc aacaacgtat cagactcttc 2521 agtttctctt gcagcaggag ttcattgtga ccgtccttct attccagcca gtttcccaga 2581 gcatcctgct tttctctcaa agaaaattgg tcaagtggaa gagcaaatag ataaagagac 2641 caagaaccca aatggggtat caagtaggga ggctaaaact gcattggatg ctgatgacag 2701 attcactttg ctgacagccc agaaaccacc tactgagtac tctaaggtag aaggcattta 2761 tacatattct ttgtctccat ccaaagtttc aggagatgat gttattgaaa aggattcccc 2821 tgaatcacca tttgaagtaa ttattgacaa agcagcattt gacaaagaat ttaaagactc 2881 atataaggag agcacagatg attttggtag ctggtctgtg cacactgata aagaatcatc 2941 cgaagacatt tcagagacta atgacaagct ttttccactg agaaataaag aggcaggacg 3001 ttacccaatg tctgcattgc tcagtaggca gttttcacac acaaatgcag cactggaaga 3061 ggtgtccaga tgcgtgaatg atatgcataa ctttactaac gaaatactga cttgggatct 3121 ggttccccaa gtgaaacaac agaccgataa atcttctgac tgcatcacaa aaactacagg 3181 acttgacatg agtgaatata attcagaaat tccagttgta aatcttaaaa ctagcactca 3241 tcagaaaact cctgtatgtt ctattgatgg gagcactccc atcactaaat caacaggtga 3301 ttgggcagaa gcatctctcc agcaagaaaa tgctattact ggaaaacctg tacctgactc 3361 tttgaattcc acaaaagaat tcagtatcaa aggtgtgcaa ggcaatatgc agaaacagga 3421 tgacacactt gcagaattac ctggatctcc acctgagaaa tgtgactctt tgggttctgg 3481 agtggccaca gtgaaagtgg ttttacctga tgaccacctg aaagatgaaa tggactggca 3541 gagctctgca ttgggagaaa tcacagaagc tgatagttct ggtgagtctg atgacacagt 3601 aatagaggac atcacagcag atacatcatt tgaaaataac aaaattcagg ctgaaaaacc 3661 tgtttccatt ccaagtgctg ttgtaaaaac aggtgaaaga gaaatcaaag agattcccag 3721 ttgtgagaga gaagaaaaaa catctaaaaa ctttgaagaa ttggtcagtg actctgagct 3781 gcatcaagat cagcctgata ttcttggaag gagtccagct agtgaggcag catgttcaaa 3841 agtacccgat acgaatgtct ccttagaaga tgtgagtgaa gttgctcctg aaaagcctat 3901 tactactgag aaccccaaac ttccttcaac agtgtctcca aatgttttta atgagacaga 3961 attctcatta aatgtgacaa catctgccta tttggagtca ttacatggga aaaatgttaa 4021 acatatagat gattcctccc cagaggacct gatagcagcc tttacagaaa ccagagataa 4081 aggaatagta gatagtgaaa gaaatgcttt taaagcaata tcagagaaga tgacagactt 4141 taaaacaact cctcctgtag aagtcttaca tgaaaatgag tccggtggtt ctgaaattaa 4201 agacattgga agcaaataca gtgaacaaag caaagaaaca aatggaagtg agcctctagg 4261 tgttttccct acccaaggta ctccagtagc atctcttgac ttagaacaag aacagctcac 4321 aattaaggct cttaaagaat taggtgaaag acaggttgag aagtcaactt ctgcacagcg 4381 tgacgcagaa ttgccttctg aagaagtact gaagcaaact ttcacatttg ctccagaatc 4441 ttggccacag agatcatatg acatcctaga acgtaatgtc aagaatggat ctgatcttgg 4501 gatttcccag aagcccatca ctatcagaga aactactagg gtagatgctg tttccagcct 4561 tagcaagact gaattggtaa aaaagcatgt cctagcaaga cttctgacag acttctcagt 4621 gcacgatctg attttctgga gagatgtgaa gaagactggg tttgtctttg gcaccacgct 4681 gatcatgctg ctttccctgg cagctttcag tgtcatcagt gtggtttctt acctcatcct 4741 ggctcttctc tctgtcacca tcagcttcag gatctacaag tccgtcatcc aagctgtaca 4801 gaagtcagaa gaaggccatc cattcaaagc ctacctggac gtagacatta ctctgtcctc 4861 agaagctttc cataattaca tgaatgctgc catggtgcac atcaacaggg ccctgaaact 4921 cattattcgt ctctttctgg tagaagatct ggttgactcc ttgaagctgg ctgtcttcat 4981 gtggctgatg acctatgttg gtgctgtttt taacggaatc acccttctaa ttcttgctga 5041 actgctcatt ttcagtgtcc cgattgtcta tgagaagtac aagacccaga ttgatcacta 5101 tgttggcatc gcccgagatc agaccaagtc aattgttgaa aagatccaag caaaactccc 5161 tggaatcgcc aaaaaaaagg cagaataagc ggccgctacg taaattccgc ccccccccct 5221 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt 5281 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg 5341 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc 5401 gtgaaggaag cagttcctct ggaagtttct tgaagacaaa caacgtctgt agcgaccctt 5461 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta 5521 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg 5581 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag 5641 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag 5701 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa 5761 acacgatgat aatatggcca caaccgtcga catgattgaa caagatggat tgcacgcagg 5821 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 5881 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 5941 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 6001 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 6061 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 6121 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 6181 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 6241 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 6301 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 6361 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 6421 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 6481 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 6541 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtcg accgcccgcc 6601 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctcga cgataaaata 6661 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg 6721 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga 6781 gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc 6841 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg 6901 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg 6961 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg 7021 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg 7081 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg 7141 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct 7201 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg 7261 actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt 7321 tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga 7381 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 7441 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 7501 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 7561 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7621 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7681 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7741 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 7801 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 7861 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7921 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7981 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 8041 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 8101 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 8161 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 8221 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 8281 ttttgcggcc ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta 8341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 8401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 8461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 8521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 8581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 8641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 8701 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 8761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 8821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 8881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 8941 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat 9001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 9061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 9121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 9181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 9241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 9301 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 9361 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cactatagga gcttggccca 9421 ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta 9481 ccgccatgtt //