LOCUS Exported 6354 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO HIS DNAJA2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6354) 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 6354) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6354 /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" protein_bind 820..838 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 841..859 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 869..893 /label=LNCX /note="Human CMV promoter, forward primer" CDS 996..1013 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2306..2323) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2312..2536 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2669..2688) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2820..2867 /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 2875..3897 /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 4027..4148 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4064..4083) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4118..4137 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4197..4213) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4197..4213) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4210..4232) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4221..4237 /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(4245..4275) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4290..4311 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4428..4445) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4599..5187) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4679..4698) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5358..6218) /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 5981..6000 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6219..6323) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttctgcaga ccatgcatca tcaccatcac catggatcca 1021 ccatggctaa cgtggctgac acgaagctgt acgacatcct gggcgtcccg cccggcgcca 1081 gcgagaacga gctgaagaag gcatacagaa agttagccaa ggaatatcat cctgataaga 1141 atccaaatgc aggagacaaa tttaaagaaa taagttttgc atatgaagta ctatcaaatc 1201 ctgagaagcg tgagttatat gacagatacg gagagcaagg tcttcgggaa ggcagcggcg 1261 gaggtggtgg catggatgat attttctctc acatttttgg tgggggattg ttcggcttca 1321 tgggcaatca gagtagaagt cgaaatggca gaagaagagg agaggacatg atgcatccac 1381 tcaaagtatc tttagaagat ctgtataatg gcaagacaac caaactacaa cttagcaaga 1441 atgtgctctg tagtgcatgc agtggccaag gcggaaagtc tggagctgtc caaaagtgta 1501 gtgcttgtcg aggtcgaggt gtgcgcatca tgatcagaca gctggctcca gggatggtac 1561 aacagatgca gtctgtgtgc tctgattgta atggagaagg agaggtaatt aatgaaaaag 1621 accgctgtaa aaaatgtgaa gggaagaagg tgattaaaga agtcaagatt cttgaagtcc 1681 acgtagacaa aggcatgaaa catggacaga gaattacatt cactggggaa gcagaccagg 1741 ccccaggagt ggaacccgga gacattgttc ttttgctaca ggagaaagaa catgaggtat 1801 ttcagagaga tgggaatgat ttgcacatga catataaaat aggacttgtt gaagctctat 1861 gtggatttca gttcacattt aagcaccttg atggacgtca gattgtggtg aaataccccc 1921 ctggcaaagt aattgaacca gggtgtgttc gtgtagttcg aggtgaaggg atgccgcagt 1981 atcgtaatcc ctttgaaaaa ggtgatcttt acataaagtt tgatgtgcag tttcctgaaa 2041 acaactggat caacccagac aagctttctg aactagaaga tcttctgcca tctagaccgg 2101 aagttcctaa cataattgga gaaacagagg aggtagagct tcaggaattt gatagcactc 2161 gaggctcagg aggtggtcag aggcgtgaag cctataatga tagctctgat gaagaaagca 2221 gcagccatca tggacctgga gtgcagtgtg cccatcagta agcggccgct cgagtctaga 2281 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt 2341 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2401 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt 2461 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 2521 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc 2581 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 2641 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 2701 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 2761 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag 2821 aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag 2881 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc 2941 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg 3001 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt 3061 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc 3121 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg 3181 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct 3241 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa 3301 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa 3361 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt 3421 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat 3481 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg 3541 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag 3601 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg 3661 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc 3721 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact 3781 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa 3841 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac 3901 gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 3961 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 4021 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 4081 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 4141 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca 4201 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 4261 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 4321 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 4381 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 4441 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 4501 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4561 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4621 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 4681 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 4741 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 4801 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 4861 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 4921 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 4981 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 5041 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5101 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5161 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5221 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 5281 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 5341 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 5401 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 5461 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 5521 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 5581 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 5641 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 5701 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 5761 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 5821 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 5881 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 5941 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 6001 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 6061 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 6121 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 6181 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 6241 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 6301 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //