LOCUS Exported 8296 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of human Bip with Myc tag near the C-terminus, retaining the C-terminal KDEL ER localization sequence. ACCESSION . VERSION . KEYWORDS pCMV BiP-Myc-KDEL-wt SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8296) AUTHORS Shen J, Chen X, Hendershot L, Prywes R TITLE ER stress regulation of ATF6 localization by dissociation of BiP/GRP78 binding and unmasking of Golgi localization signals. JOURNAL Dev Cell. 2002 Jul . 3(1):99-111. PUBMED 12110171 REFERENCE 2 (bases 1 to 8296) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8296 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 178..557 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 558..761 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 712..732 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 2776..2805 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2839..2904 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal 2915..3537 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3114..3133) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(3561..3581) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3579..3876 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind 3808..3827 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3930..4724 /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(3984..4003) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4594..4613 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5376..5510 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5413..5432) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5467..5486 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5515..5534) /label=T7 /note="T7 promoter, forward primer" promoter complement(5516..5534) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5548..5564) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5548..5564) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5561..5583) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5572..5588 /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(5596..5626) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5641..5662 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5779..5796) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5950..6538) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6030..6049) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6709..7569) /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 7332..7351 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7570..7674) /gene="bla" /label=AmpR promoter rep_origin 7701..8156 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7788..7807) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7998..8019 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8282..8 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8296..17 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" ORIGIN 1 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca 61 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt 121 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac 181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 421 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 481 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag 541 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt 601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 661 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg 721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga 781 attaagcttg ctggcaagat gaagctctcc ctggtggccg cgatgctgct gctgctcagc 841 gcggcgcggg ccgaggagga ggacaagaag gaggacgtgg gcacggtggt cggcatcgac 901 ctggggacca cctactcctg cgtcggcgtg ttcaagaacg gccgcgtgga gatcatcgcc 961 aacgatcagg gcaaccgcat cacgccgtcc tatgtcgcct tcactcctga aggggaacgt 1021 ctgattggcg atgccgccaa gaaccagctc acctccaacc ccgagaacac ggtctttgac 1081 gccaagcggc tcatcggccg cacgtggaat gacccgtctg tgcagcagga catcaagttc 1141 ttgccgttca aggtggttga aaagaaaact aaaccataca ttcaagttga tattggaggt 1201 gggcaaacaa agacatttgc tcctgaagaa atttctgcca tggttctcac taaaatgaaa 1261 gaaaccgctg aggcttattt gggaaagaag gttacccatg cagttgttac tgtaccagcc 1321 tattttaatg atgcccaacg ccaagcaacc aaagacgctg gaactattgc tggcctaaat 1381 gttatgagga tcatcaacga gcctacggca gctgctattg cttatggcct ggataagagg 1441 gagggggaga agaacatcct ggtgtttgac ctgggtggcg gaaccttcga tgtgtctctt 1501 ctcaccattg acaatggtgt cttcgaagtt gtggccacta atggagatac tcatctgggt 1561 ggagaagact ttgaccagcg tgtcatggaa cacttcatca aactgtacaa aaagaagacg 1621 ggcaaagatg tcaggaaaga caatagagct gtgcagaaac tccggcgcga ggtagaaaag 1681 gccaaacggg ccctgtcttc tcagcatcaa gcaagaattg aaattgagtc cttctatgaa 1741 ggagaagact tttctgagac cctgactcgg gccaaatttg aagagctcaa catggatctg 1801 ttccggtcta ctatgaagcc cgtccagaaa gtgttggaag attctgattt gaagaagtct 1861 gatattgatg aaattgttct tgttggtggc tcgactcgaa ttccaaagat tcagcaactg 1921 gttaaagagt tcttcaatgg caaggaacca tcccgtggca taaacccaga tgaagctgta 1981 gcgtatggtg ctgctgtcca ggctggtgtg ctctctggtg atcaagatac aggtgacctg 2041 gtactgcttg atgtatgtcc ccttacactt ggtattgaaa ctgtgggagg tgtcatgacc 2101 aaactgattc caaggaacac agtggtgcct accaagaagt ctcagatctt ttctacagct 2161 tctgataatc aaccaactgt tacaatcaag gtctatgaag gtgaaagacc cctgacaaaa 2221 gacaatcatc ttctgggtac atttgatctg actggaattc ctcctgctcc tcgtggggtc 2281 ccacagattg aagtcacctt tgagatagat gtgaatggta ttcttcgagt gacagctgaa 2341 gacaagggta cagggaacaa aaataagatc acaatcacca atgaccagaa tcgcctgaca 2401 cctgaagaaa tcgaaaggat ggttaatgat gctgagaagt ttgctgagga agacaaaaag 2461 ctcaaggagc gcattgatac tagaaatgag ttggaaagct atgcctattc tctaaagaat 2521 cagattggag ataaagaaaa gctgggaggt aaactttcct ctgaagataa ggagaccatg 2581 gaaaaagctg tagaagaaaa gattgaatgg ctggaaagcc accaagatgc tgacattgaa 2641 gacttcaaag ctaagaagaa ggaactggaa gaaattgttc aaccaattat cagcaaactc 2701 tatggaagtg caggccctcc cccaactggt gaagaggata cagcagaaaa agatgagttg 2761 gctagagggc ccttcgaaca aaaactcatc tcagaagagg atctgaagga cgagctgtag 2821 tctagaggat cccgggctga ctacaaagac catgacggtg attataaaga tcatgacatc 2881 gactacaagg atgacgatga caagtagtga tcccgggtgg catccctgtg acccctcccc 2941 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa 3001 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg 3061 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg 3121 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt 3181 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag 3241 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt 3301 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca 3361 ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat accagcagga 3421 ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg 3481 cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg 3541 tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc 3601 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa 3661 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3721 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 3781 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 3841 tgagctattc cagaagtagt gaggaggctt ttttggagga attgatcagc ttgggatctg 3901 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3961 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 4021 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 4081 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 4141 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 4201 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 4261 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 4321 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 4381 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 4441 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 4501 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 4561 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4621 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4681 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4741 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 4801 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 4861 gcgcggggat ctcatgctgg agttcttcgc ccaccccggg ctcgatcccc tcgcgagttg 4921 gttcagctgc tgcctgaggc tggacgacct cgcggagttc taccggcagt gcaaatccgt 4981 cggcatccag gaaaccagca gcggctatcc gcgcatccat gcccccgaac tgcaggagtg 5041 gggaggcacg atggccgctt tggtcgaccc ggacgggacg ctcctgcgcc tgatacagaa 5101 cgaattgctt gcaggcatct catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg 5161 tggatggagc gctggcgcct gctgcgcgac ggcgagctgc tcaccaccca ctcgccaagc 5221 tggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccg atcataatca 5281 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga 5341 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg 5401 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 5461 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aattccctat 5521 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 5581 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 5641 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 5701 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 5761 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 5821 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 5881 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 5941 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6001 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6061 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6121 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6181 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6241 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6301 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6361 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 6421 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 6481 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 6541 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6601 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 6661 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6721 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6781 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6841 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6901 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6961 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7021 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7081 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7141 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7201 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7261 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7321 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7381 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7441 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7501 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 7561 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 7621 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 7681 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 7741 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 7801 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 7861 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 7921 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 7981 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 8041 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 8101 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat 8161 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 8221 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt 8281 tcccagtcac gacgtt //