LOCUS Exported 7334 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3.1(+)-GRP78/BiP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7334) AUTHORS Werstuck GH, Lentz SR, Dayal S, Hossain GS, Sood SK, Shi YY, Zhou J, Maeda N, Krisans SK, Malinow MR, Austin RC TITLE Homocysteine-induced endoplasmic reticulum stress causes dysregulation of the cholesterol and triglyceride biosynthetic pathways. JOURNAL J Clin Invest. 2001 May;107(10):1263-73. PUBMED 11375416 REFERENCE 2 (bases 1 to 7334) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7334 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 865..884 /label=T7 /note="T7 promoter, forward primer" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2927..2944) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2933..3157 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3203..3631 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3290..3309) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3500..3521 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3640..3660) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3645..3974 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3825..3960 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3887..3906 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4041..4835 /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(4095..4114) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4705..4724 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5009..5130 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5046..5065) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5100..5119 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5179..5195) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5179..5195) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5192..5214) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5203..5219 /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(5227..5257) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5272..5293 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5410..5427) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5581..6169) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5661..5680) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6340..7200) /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 6963..6982 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7201..7305) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta 901 gcgtttaaac ttaagcttgc caccatgaag ctctccctgg tggccgcgat gctgctgctg 961 ctcagcgcgg cgcgggccga ggaggaggac aagaaggagg acgtgggcac ggtggtcggc 1021 atcgacctgg ggaccaccta ctcctgcgtc ggcgtgttca agaacggccg cgtggagatc 1081 atcgccaacg atcagggcaa ccgcatcacg ccgtcctatg tcgccttcac tcctgaaggg 1141 gaacgtctga ttggcgatgc cgccaagaac cagctcacct ccaaccccga gaacacggtc 1201 tttgacgcca agcggctcat cggccgcacg tggaatgacc cgtctgtgca gcaggacatc 1261 aagttcttgc cgttcaaggt ggttgaaaag aaaactaaac catacattca agttgatatt 1321 ggaggtgggc aaacaaagac atttgctcct gaagaaattt ctgccatggt tctcactaaa 1381 atgaaagaaa ccgctgaggc ttatttggga aagaaggtta cccatgcagt tgttactgta 1441 ccagcctatt ttaatgatgc ccaacgccaa gcaaccaaag acgctggaac tattgctggc 1501 ctaaatgtta tgaggatcat caacgagcct acggcagctg ctattgctta tggcctggat 1561 aagagggagg gggagaagaa catcctggtg tttgacctgg gtggcggaac cttcgatgtg 1621 tctcttctca ccattggcaa tggtgtcttc gaagttgtgg ccactaatgg agatactcat 1681 ctgggtggag aagactttga ccagcgtgtc atggaacact tcatcaaact gtacaaaaag 1741 aagacgggca aagatgtcag gaaagacaat agagctgtgc agaaactccg gcgcgaggta 1801 gaaaaggcca aacgggccct gtcttctcag catcaagcaa gaattgaaat tgagtccttc 1861 tatgaaggag aagacttttc tgagaccctg actcgggcca aatttgaaga gctcaacatg 1921 gatctgttcc ggtctactat gaagcccgtc cagaaagtgt tggaagattc tgatttgaag 1981 aagtctgata ttgatgaaat tgttcttgtt ggtggctcga ctcgaattcc aaagattcag 2041 caactggtta aagagttctt caatggcaag gaaccatccc gtggcataaa cccagatgaa 2101 gctgtagcgt atggtgctgc tgtccaggct ggtgtgctct ctggtgatca agatacaggt 2161 gacctggtac tgcttgatgt atgtcccctt acacttggta ttgaaactgt gggaggtgtc 2221 atgaccaaac tgattccaag gaacacagtg gtgcctacca agaagtctca gatcttttct 2281 acagcttctg ataatcaacc aactgttaca atcaaggtct atgaaggtga aagacccctg 2341 acaaaagaca atcatcttct gggtacattt gatctgactg gaattcctcc tgctcctcgt 2401 ggggtcccac agattgaagt cacctttgag atagatgtga atggtattct tcgagtgaca 2461 gctgaagaca agggtacagg gaacaaaaat aagatcacaa tcaccaatga ccagaatcgc 2521 ctgacacctg aagaaatcga aaggatggtt aatgatgctg agaagtttgc tgaggaagac 2581 aaaaagctca aggagcgcat tgatactaga aatgagttgg aaagctatgc ctattctcta 2641 aagaatcaga ttggagataa agaaaagctg ggaggtaaac tttcctctga agataaggag 2701 accatggaaa aagctgtaga agaaaagatt gaatggctgg aaagccacca agatgctgac 2761 attgaagact tcaaagctaa gaagaaggaa ctggaagaaa ttgttcaacc aattatcagc 2821 aaactctatg gaagtgcagg ccctccccca actggtgaag aggatacagc agaaaaagat 2881 gagttgtagc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct 2941 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 3001 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 3061 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 3121 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 3181 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 3241 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 3301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 3361 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 3421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 3481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 3541 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 3601 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3721 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3781 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3901 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 3961 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga 4021 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 4081 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 4141 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 4201 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg 4261 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 4321 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 4381 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 4441 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 4501 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 4561 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4621 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 4681 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 4741 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4801 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 4861 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4921 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4981 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 5041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 5101 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta 5161 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5221 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 5281 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5341 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5401 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5461 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5521 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5581 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 5641 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5701 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 5761 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 5821 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 5881 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 5941 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6001 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 6061 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6121 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6181 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6241 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6301 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6361 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6421 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6481 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6541 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6601 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 6661 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 6721 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 6781 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 6841 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 6901 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 6961 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7021 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7081 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7141 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7201 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7261 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7321 agtgccacct gacg //