LOCUS Exported 7672 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO HIS HSPH3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7672) 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 7672) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7672 /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(3624..3641) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3630..3854 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3987..4006) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 4138..4185 /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 4193..5215 /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 5345..5466 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5382..5401) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5436..5455 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5515..5531) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5515..5531) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5528..5550) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5539..5555 /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(5563..5593) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5608..5629 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5746..5763) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5917..6505) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5997..6016) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6676..7536) /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 7299..7318 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7537..7641) /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 ctagtccagt gtggtggaat tctgcagata tcaccatgtc tgtggttggc attgacctcg 1081 gctttctcaa ctgctacatt gctgtcgcga gaagtggcgg catcgagacc atcgccaatg 1141 agtacagcga caggtgtacc ccggcctgta tatcattggg atcaagaact cgagccattg 1201 gaaatgcagc aaagagccag atagtcacga acgtaagaaa tacaattcat ggcttcaaaa 1261 agcttcatgg gcgatcattt gatgatccca ttgtgcaaac tgaaaggatc aggcttccct 1321 atgaactgca gaaaatgcct aatggaagtg caggagttaa ggtgcggtac ttagaggaag 1381 agagaccttt tgcaattgag caagttactg gaatgctgtt agccaagctt aaagagactt 1441 cagaaaatgc tttgaagaaa ccagtggctg actgtgtgat ttcaattcct agctttttta 1501 ctgatgccga gagaagatct gtgatggctg cagcccaggt tgcaggctta aattgtttaa 1561 ggttgatgaa tgaaactact gcagttgcac tggcgtatgg aatttataaa caggatcttc 1621 ccccattaga tgagaaacca agaaatgtag tatttattga tatgggacat tctgcctatc 1681 aggtcttggt ttgtgctttt aacaaaggaa aacttaaagt cttggctact acctttgatc 1741 catatttggg tggcaggaac tttgatgagg ctttagtaga ctacttctgt gatgagttca 1801 agaccaaata taagataaat gtgaaagaaa actctcgggc cttgttgcgt ttatatcagg 1861 aatgtgaaaa actaaagaag ctaatgagtg caaatgcatc agatcttcca ttgaacattg 1921 agtgtttcat gaatgacctt gatgtttcta gtaaaatgaa cagggctcaa tttgaacaac 1981 tgtgtgcttc ccttttggcc agggttgaac cacctttaaa agcagtaatg gaacaagcta 2041 acttacaacg tgaagacatt agtagtatag aaattgtagg aggagcaaca cgaattcctg 2101 cagtgaaaga acaaatcact aaattctttc ttaaagacat aagtaccaca ttaaatgctg 2161 atgaagctgt tgcaagagga tgtgcgttac agtgtgcgat tctctcacca gcatttaaag 2221 tgcgtgaatt ttccataaca gaccttgttc cctattcaat cacattaagg tggaagacct 2281 cttttgaaga tggaagtggg gaatgtgaag ttttctgtaa gaaccatcct gccccattct 2341 caaaagtcat tactttccac aagaaggaac catttgaact agaagcattt tatactaatt 2401 tacatgaagt gccttatcct gatgcaagaa ttgggagctt cactattcag aatgtttttc 2461 cacagtctga tggtgatagt tccaaagtga aggttaaagt tcgtgttaac atccatggaa 2521 tcttcagtgt ggctagcgca tcagtaattg agaagcaaaa tttggaaggc gatcacagtg 2581 atgctccaat ggagacagaa acttcattta aaaatgaaaa caaagataat atggataaaa 2641 tgcaggttga tcaagaagaa gggcatcaaa aatgtcatgc tgaacacact ccagaagagg 2701 aaattgatca tacaggagcc aaaacaaagt cagctgtctc agacaaacaa gaccgattaa 2761 atcagacact taaaaaagga aaagtcaaaa gtattgatct accgatccag agtagcctat 2821 gtagacaact aggccaagat cttctcaaca gctacattga aaatgagggg aagatgatca 2881 tgcaagataa gttagagaaa gaaagaaatg atgctaagaa tgccgttgaa gaatatgtat 2941 atgattttag agacaggctg ggcactgtct atgaaaaatt catcactcca gaagacttga 3001 gtaaactgtc tgcagtattg gaagacacag aaaattggct ttatgaagac ggagaggacc 3061 aacctaaaca agtttatgtg gataagcttc aagaactaaa gaaatacggc cagcctattc 3121 aaatgaagta catggagcat gaagagagac caaaagcctt aaatgacttg ggaaaaaaga 3181 tccaacttgt catgaaagtg atagaagctt atagaaacaa ggatgaaaga tatgatcatc 3241 tggatcctac tgaaatggaa aaggttgaaa aatgtatcag tgatgccatg agttggctga 3301 atagtaagat gaatgcacag aacaaactaa gtctcactca agatcctgtg gtaaaagttt 3361 cagaaatagt agcaaagtca aaggaactgg ataatttctg taaccccatc atttacaagc 3421 ccaaaccaaa agcagaagtt cctgaagaca aaccaaaagc taatagtgaa cacaatggcc 3481 caatggatgg acagagtgga actgaaacta aatcagattc aacaaaagac agctcacagc 3541 atactaaatc ctctggagag atggaagtgg actaagtctg cggccgctcg agtctagagg 3601 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt 3661 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta 3721 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg 3781 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc 3841 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca 3901 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 3961 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 4021 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 4081 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa 4141 gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc 4201 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga 4261 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg 4321 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta 4381 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag 4441 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc 4501 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc 4561 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata 4621 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac 4681 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg 4741 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt 4801 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga 4861 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca 4921 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc 4981 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga 5041 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt 5101 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt 5161 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt 5221 actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 5281 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 5341 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 5401 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 5461 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 5521 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 5581 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 5641 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 5701 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 5761 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 5821 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 5881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 5941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 6001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 6061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 6121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 6181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 6241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 6301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 6361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 6421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 6481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 6541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 6601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 6661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 6721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 6781 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 6841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 6901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 6961 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 7021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 7081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 7141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 7201 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 7261 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 7321 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 7381 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 7441 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 7501 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 7561 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 7621 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //