LOCUS Exported 7091 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS AIRAP 10: PSMD14.TEV.GST.V1.pCDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7091) AUTHORS Stanhill A, Haynes CM, Zhang Y, Min G, Steele MC, Kalinina J, Martinez E, Pickart CM, Kong XP, Ron D TITLE An arsenite-inducible 19S regulatory particle-associated protein adapts proteasomes to proteotoxicity. JOURNAL Mol Cell. 2006 Sep 15. 23(6):875-85. PUBMED 16973439 REFERENCE 2 (bases 1 to 7091) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7091 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1895..1915 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 1922..2575 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 2533..2555 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 2582..2599 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" promoter complement(2643..2661) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2644..2661) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2681..2698) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2687..2911 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2957..3385 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3044..3063) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3254..3275 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3394..3414) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3399..3729 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3580..3715 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3642..3661 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3796..4590 /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(3850..3869) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4460..4479 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4764..4885 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4801..4820) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4855..4874 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4934..4950) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4934..4950) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4947..4969) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4958..4974 /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(4982..5012) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5027..5048 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5165..5182) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5336..5924) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5416..5435) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6095..6955) /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 6718..6737 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6956..7060) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc 901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttccagttac tgaacagaaa 961 tatggacaga cttcttagac ttggaggagg tatgcctgga ctgggccagg gcccacctac 1021 agatgctcct gccgtggaca cagcagaaca agtttatatc tcttccttgg ccttgctaaa 1081 gatgttaaaa catggtcgtg ctggagtgcc tatggaagtt atgggtctaa tgcttggtga 1141 atttgttgat gattacaccg tcagagtgat tgatgtgttt gctatgccac agtcaggaac 1201 tggtgtcagt gttgaagcag ttgatccagt gttccaagcc aaaatgttgg atatgctgaa 1261 acaaacagga aggcccgaga tggttgttgg ttggtatcac agtcaccctg gctttggctg 1321 ttggctttct ggtgtggata tcaacactca gcagagcttt gaagccttgt cggagagagc 1381 tgtggcagtg gttgtggatc ccattcagag tgtaaaagga aaggttgtta ttgatgcctt 1441 cagactgatc aatgctaata tgatggtctt aggacatgaa ccaagacaaa cgacttcaaa 1501 tctgggccac ttaaacaagc catctatcca ggcattaatt cacggactaa acagacatta 1561 ttactccatc actattaatt accggaaaaa tgaactggaa cagaagatgc tgttaaattt 1621 gcataagaag agttggatgg aaggattgac acttcaggac tacagtgaac actgtaaaca 1681 caatgaatcg gtggtaaaag aaatgttgga attagccaag aattataata aggctgtgga 1741 agaagaagat aagatgacac ctgaacagct ggcaataaag aatgttggca agcaggatcc 1801 caaacgtcat ttggaagaac atgtggatgt gcttatgact tcaaatattg tccagtgttt 1861 ggccgcaatg ttggatactg ttgtatttaa actcgagaac ctgtatttcc agggtaccga 1921 gatgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct 1981 tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa 2041 atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga 2101 tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa 2161 catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt 2221 ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt 2281 tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa 2341 aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga 2401 tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa 2461 aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc 2521 atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga 2581 tctggttccg cgtggatccc cgggaatttc cggtggtggt ggtggaatct agagggccct 2641 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt 2701 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 2761 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 2821 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 2881 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 2941 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3001 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3061 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct 3121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 3241 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3301 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg 3361 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3421 agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 3541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 3601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 3661 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 3721 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga 3781 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 3841 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 3901 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 3961 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 4021 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 4081 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 4141 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 4201 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 4261 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 4321 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 4381 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 4441 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 4501 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 4561 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 4621 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 4681 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 4741 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 4801 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 4861 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 4921 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4981 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 5041 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 5101 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 5161 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 5221 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 5281 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 5341 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 5401 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 5461 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 5521 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 5581 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 5641 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 5701 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 5761 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 5821 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 5881 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5941 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 6001 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 6061 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 6121 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 6181 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 6241 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 6301 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 6361 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 6421 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 6481 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 6541 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6601 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6661 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6721 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6781 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6841 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6901 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6961 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 7021 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 7081 cacctgacgt c //