LOCUS Exported 7654 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO HSPH3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7654) 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 7654) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7654 /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" primer_bind complement(3606..3623) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3612..3836 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3969..3988) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 4120..4167 /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 4175..5197 /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 5327..5448 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5364..5383) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5418..5437 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5497..5513) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5497..5513) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5510..5532) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5521..5537 /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(5545..5575) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5590..5611 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5728..5745) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5899..6487) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5979..5998) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6658..7518) /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 7281..7300 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7519..7623) /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 cttggtaccg agctcggatc cactagtcca gtgtggtgga 1021 attctgcaga tatcaccatg tctgtggttg gcattgacct cggctttctc aactgctaca 1081 ttgctgtcgc gagaagtggc ggcatcgaga ccatcgccaa tgagtacagc gacaggtgta 1141 ccccggcctg tatatcattg ggatcaagaa ctcgagccat tggaaatgca gcaaagagcc 1201 agatagtcac gaacgtaaga aatacaattc atggcttcaa aaagcttcat gggcgatcat 1261 ttgatgatcc cattgtgcaa actgaaagga tcaggcttcc ctatgaactg cagaaaatgc 1321 ctaatggaag tgcaggagtt aaggtgcggt acttagagga agagagacct tttgcaattg 1381 agcaagttac tggaatgctg ttagccaagc ttaaagagac ttcagaaaat gctttgaaga 1441 aaccagtggc tgactgtgtg atttcaattc ctagcttttt tactgatgcc gagagaagat 1501 ctgtgatggc tgcagcccag gttgcaggct taaattgttt aaggttgatg aatgaaacta 1561 ctgcagttgc actggcgtat ggaatttata aacaggatct tcccccatta gatgagaaac 1621 caagaaatgt agtatttatt gatatgggac attctgccta tcaggtcttg gtttgtgctt 1681 ttaacaaagg aaaacttaaa gtcttggcta ctacctttga tccatatttg ggtggcagga 1741 actttgatga ggctttagta gactacttct gtgatgagtt caagaccaaa tataagataa 1801 atgtgaaaga aaactctcgg gccttgttgc gtttatatca ggaatgtgaa aaactaaaga 1861 agctaatgag tgcaaatgca tcagatcttc cattgaacat tgagtgtttc atgaatgacc 1921 ttgatgtttc tagtaaaatg aacagggctc aatttgaaca actgtgtgct tcccttttgg 1981 ccagggttga accaccttta aaagcagtaa tggaacaagc taacttacaa cgtgaagaca 2041 ttagtagtat agaaattgta ggaggagcaa cacgaattcc tgcagtgaaa gaacaaatca 2101 ctaaattctt tcttaaagac ataagtacca cattaaatgc tgatgaagct gttgcaagag 2161 gatgtgcgtt acagtgtgcg attctctcac cagcatttaa agtgcgtgaa ttttccataa 2221 cagaccttgt tccctattca atcacattaa ggtggaagac ctcttttgaa gatggaagtg 2281 gggaatgtga agttttctgt aagaaccatc ctgccccatt ctcaaaagtc attactttcc 2341 acaagaagga accatttgaa ctagaagcat tttatactaa tttacatgaa gtgccttatc 2401 ctgatgcaag aattgggagc ttcactattc agaatgtttt tccacagtct gatggtgata 2461 gttccaaagt gaaggttaaa gttcgtgtta acatccatgg aatcttcagt gtggctagcg 2521 catcagtaat tgagaagcaa aatttggaag gcgatcacag tgatgctcca atggagacag 2581 aaacttcatt taaaaatgaa aacaaagata atatggataa aatgcaggtt gatcaagaag 2641 aagggcatca aaaatgtcat gctgaacaca ctccagaaga ggaaattgat catacaggag 2701 ccaaaacaaa gtcagctgtc tcagacaaac aagaccgatt aaatcagaca cttaaaaaag 2761 gaaaagtcaa aagtattgat ctaccgatcc agagtagcct atgtagacaa ctaggccaag 2821 atcttctcaa cagctacatt gaaaatgagg ggaagatgat catgcaagat aagttagaga 2881 aagaaagaaa tgatgctaag aatgccgttg aagaatatgt atatgatttt agagacaggc 2941 tgggcactgt ctatgaaaaa ttcatcactc cagaagactt gagtaaactg tctgcagtat 3001 tggaagacac agaaaattgg ctttatgaag acggagagga ccaacctaaa caagtttatg 3061 tggataagct tcaagaacta aagaaatacg gccagcctat tcaaatgaag tacatggagc 3121 atgaagagag accaaaagcc ttaaatgact tgggaaaaaa gatccaactt gtcatgaaag 3181 tgatagaagc ttatagaaac aaggatgaaa gatatgatca tctggatcct actgaaatgg 3241 aaaaggttga aaaatgtatc agtgatgcca tgagttggct gaatagtaag atgaatgcac 3301 agaacaaact aagtctcact caagatcctg tggtaaaagt ttcagaaata gtagcaaagt 3361 caaaggaact ggataatttc tgtaacccca tcatttacaa gcccaaacca aaagcagaag 3421 ttcctgaaga caaaccaaaa gctaatagtg aacacaatgg cccaatggat ggacagagtg 3481 gaactgaaac taaatcagat tcaacaaaag acagctcaca gcatactaaa tcctctggag 3541 agatggaagt ggactaagtc tgcggccgct cgagtctaga gggcccgttt aaacccgctg 3601 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 3661 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 3721 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 3781 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 3841 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 4081 ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc 4141 ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc 4201 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga 4261 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt 4321 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc 4381 cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg 4441 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc 4501 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac 4561 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt 4621 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt 4681 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga 4741 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg 4801 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc 4861 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga 4921 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg 4981 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca 5041 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc 5101 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa 5161 ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc 5221 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 5281 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc 5341 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 5401 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 5461 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 5521 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 5581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 5701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 5761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 5821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 5881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 5941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 6001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 6061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 6121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 6361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 6841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 6901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 7021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 7081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 7141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 7561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7621 gcgcacattt ccccgaaaag tgccacctga cgtc //