LOCUS Exported 5844 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO HIS DNAJB3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5844) 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 5844) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5844 /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(1796..1813) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1802..2026 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2159..2178) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2310..2357 /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 2365..3387 /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 3517..3638 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3554..3573) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3608..3627 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3687..3703) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3687..3703) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3700..3722) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3711..3727 /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(3735..3765) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3780..3801 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3918..3935) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4089..4677) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4169..4188) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4848..5708) /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 5471..5490 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5709..5813) /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 ccatggtgga ctactacgag gtgctgggcg tgccgcggca ggcctcagcc gaggccatcc 1081 gcaaggcgta ccgcaagctt gcgctcaagt ggcacccgga caagaacccc gagcacaagg 1141 aggaggccga gaggcggttc aagcaggtgg cgcaggccta cgaggtctta tcggacgtcc 1201 gcaagcgcga ggtgtacgac cgctgcggcg aagtgggcga ggtgggcgga ggcggcgcgg 1261 cgggcagccc gttccacgat gccttccagt acgtcttctc cttccgagac cccgcggagg 1321 tcttcaggga gttcttcggg ggccacgacc ctttctcctt cgacttcttc gggggagacc 1381 cccttgagaa cttttttggg gatcggagga gcacccgtgg aagcagaagc cgaggggctg 1441 tacccttctc tacctccttt accgaattcc caggatttgg gggtggcttc gcttcgttag 1501 atactggatt cacatccttc ggttctccag gaaactcggg cctttcttct ttctccatgt 1561 cttgcggcgg tggggcggca ggcaactaca agtcggtgtc aacctccacc gaaataatta 1621 atggcaaaaa aatcaccacc aagagaatcg ttgagaatgg tcaagaaagg gtggaagtgg 1681 aagaagatgg agagttaaag tccctgataa ttaatggcag agagcagctg cttcgcatca 1741 atactcagta agcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg 1801 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 1861 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 1921 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 1981 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 2041 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 2101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 2221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 2281 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta 2341 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag 2401 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa 2461 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc 2521 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg 2581 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc 2641 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag 2701 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc 2761 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg 2821 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc 2881 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac 2941 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg 3001 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc 3061 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat 3121 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa 3181 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc 3241 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc 3301 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc 3361 agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc 3421 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 3481 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 3541 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 3601 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 3661 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 3721 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 3781 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 3841 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 3901 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 3961 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 4021 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 4081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 4141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 4201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 4261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 4321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 4381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 4441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 4501 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 4561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 4621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 4681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 4741 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 4801 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 4861 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 4921 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 4981 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 5041 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 5101 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 5161 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 5221 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 5281 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 5341 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 5401 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 5461 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 5521 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 5581 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 5641 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 5701 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 5761 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 5821 ccccgaaaag tgccacctga cgtc //