LOCUS Exported 6831 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO GFP DNAJB1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6831) AUTHORS Hageman J, Kampinga HH TITLE Computational analysis of the human HSPH/HSPA/DNAJ family and cloning of a human HSPH/HSPA/DNAJ expression library. JOURNAL Cell Stress Chaperones. 2008 Aug 7. ():. PUBMED 18686016 REFERENCE 2 (bases 1 to 6831) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6831 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 6..15 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 12..728 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 12..728 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(57..78) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(318..337) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 665..686 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1805..1822) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1811..2035 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2168..2187) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2319..2366 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 2374..3396 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 3526..3647 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3563..3582) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3617..3636 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3696..3712) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3696..3712) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3709..3731) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3720..3736 /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(3744..3774) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3789..3810 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3927..3944) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4098..4686) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4178..4197) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4857..5717) /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 5480..5499 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5718..5822) /gene="bla" /label=AmpR promoter primer_bind complement(5897..5916) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 6088..6467 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6468..6671 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6622..6642 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 6673..6691 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 6694..6712 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 6722..6746 /label=LNCX /note="Human CMV promoter, forward primer" ORIGIN 1 agcttgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg 61 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 121 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc 181 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg 241 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 301 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 361 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 421 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca 481 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg 541 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 601 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg 661 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc 721 tgtacaaggg atccaccatg ggtaaagact actaccagac gttgggcctg gcccgcggcg 781 cgtcggacga ggagatcaag cgggcctacc gccgccaggc gctgcgctac cacccggaca 841 agaacaagga gcccggcgcc gaggagaagt tcaaggagat cgctgaggcc tacgacgtgc 901 tcagcgaccc gcgcaagcgc gagatcttcg accgctacgg ggaggaaggc ctaaagggga 961 gtggccccag tggcggtagc ggcggtggtg ccaatggtac ctctttcagc tacacattcc 1021 atggagaccc tcatgccatg tttgctgagt tcttcggtgg cagaaatccc tttgacacct 1081 tttttgggca gcggaacggg gaggaaggca tggacattga tgacccattc tctggcttcc 1141 ctatgggcat gggtggcttc accaacgtga actttggccg ctcccgctct gcccaagagc 1201 ccgcccgaaa gaagcaagat cccccagtca cccacgacct tcgagtctcc cttgaagaga 1261 tctacagcgg ctgtaccaag aagatgaaaa tctcccacaa gcggctaaac cccgacggaa 1321 agagcattcg aaacgaagac aaaatattga ccatcgaagt gaagaagggg tggaaagaag 1381 gaaccaaaat cactttcccc aaggaaggag accagacctc caacaacatt ccagctgata 1441 tcgtctttgt tttaaaggac aagccccaca atatctttaa gagagatggc tctgatgtca 1501 tttatcctgc caggatcagc ctccgggagg ctctgtgtgg ctgcacagtg aacgtcccca 1561 ctctggacgg caggacgata cccgtcgtat tcaaagatgt tatcaggcct ggcatgcggc 1621 gaaaagttcc tggagaaggc ctccccctcc ccaaaacacc cgagaaacgt ggggacctca 1681 ttattgagtt tgaagtgatc ttccccgaaa ggattcccca gacatcaaga accgtacttg 1741 agcaggttct tccaatatag gcggccgctc gagtctagag ggcccgttta aacccgctga 1801 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 1861 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 1921 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 1981 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 2041 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2101 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2161 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2221 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 2281 cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt ccgaagttcc 2341 tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac cgcgacgtct 2401 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag 2461 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta 2521 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc 2581 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc 2641 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct 2701 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg 2761 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc 2821 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc 2881 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa 2941 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc 3001 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc 3061 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag 3121 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt 3181 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag 3241 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc 3301 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac 3361 cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga tttcgattcc 3421 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 3481 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca 3541 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 3601 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata 3661 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 3721 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 3781 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 3841 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 3901 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 3961 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 4021 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 4081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 4141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 4201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 4261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 4321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 4381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 4441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 4501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 4561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 4621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 4681 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 4741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 4801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 4861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 4921 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 4981 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 5041 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 5101 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 5161 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 5221 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 5281 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 5341 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 5401 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 5461 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 5521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 5581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 5641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 5701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 5761 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 5821 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc 5881 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc 5941 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc 6001 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc 6061 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat 6121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 6181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 6241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 6301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 6361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 6421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 6481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 6541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 6601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 6661 taagcagagc tctccctatc agtgatagag atctccctat cagtgataga gatcgtcgac 6721 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc 6781 atagaagaca ccgggaccga tccagcctcc ggactctagc gtttaaactt a //