LOCUS Exported 7056 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO HIS HSPA8 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7056) 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 7056) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7056 /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(3008..3025) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3014..3238 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3371..3390) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3522..3569 /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 3577..4599 /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 4729..4850 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4766..4785) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4820..4839 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4899..4915) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4899..4915) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4912..4934) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4923..4939 /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(4947..4977) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4992..5013 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5130..5147) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5301..5889) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5381..5400) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6060..6920) /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 6683..6702 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6921..7025) /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 ccatgtccaa gggacctgca gttggtattg atcttggcac cacctactct tgtgtgggtg 1081 ttttccagca cggaaaagtc gagataattg ccaatgatca gggaaaccga accactccaa 1141 gctatgtcgc ctttacggac actgaacggt tgatcggtga tgccgcaaag aatcaagttg 1201 caatgaaccc caccaacaca gtttttgatg ccaaacgtct gattggacgc agatttgatg 1261 atgctgttgt ccagtctgat atgaaacatt ggccctttat ggtggtgaat gatgctggca 1321 ggcccaaggt ccaagtagaa tacaagggag agaccaaaag cttctatcca gaggaggtgt 1381 cttctatggt tctgacaaag atgaaggaaa ttgcagaagc ctaccttggg aagactgtta 1441 ccaatgctgt ggtcacagtg ccagcttact ttaatgactc tcagcgtcag gctaccaaag 1501 atgctggaac tattgctggt ctcaatgtac ttagaattat taatgagcca actgctgctg 1561 ctattgctta cggcttagac aaaaaggttg gagcagaaag aaacgtgctc atctttgacc 1621 tgggaggtgg cacttttgat gtgtcaatcc tcactattga ggatggaatc tttgaggtca 1681 agtctacagc tggagacacc cacttgggtg gagaagattt tgacaaccga atggtcaacc 1741 attttattgc tgagtttaag cgcaagcata agaaggacat cagtgagaac aagagagctg 1801 taagacgcct ccgtactgct tgtgaacgtg ctaagcgtac cctctcttcc agcacccagg 1861 ccagtattga gatcgattct ctctatgaag gaatcgactt ctatacctcc attacccgtg 1921 cccgatttga agaactgaat gctgacctgt tccgtggcac cctggaccca gtagagaaag 1981 cccttcgaga tgccaaacta gacaagtcac agattcatga tattgtcctg gttggtggtt 2041 ctactcgtat ccccaagatt cagaagcttc tccaagactt cttcaatgga aaagaactga 2101 ataagagcat caaccctgat gaagctgttg cttatggtgc agctgtccag gcagccatct 2161 tgtctggaga caagtctgag aatgttcaag atttgctgct cttggatgtc actcctcttt 2221 cccttggtat tgaaactgct ggtggagtca tgactgtcct catcaagcgt aataccacca 2281 ttcctaccaa gcagacacag accttcacta cctattctga caaccagcct ggtgtgctta 2341 ttcaggttta tgaaggcgag cgtgccatga caaaggataa caacctgctt ggcaagtttg 2401 aactcacagg catacctcct gcaccccgag gtgttcctca gattgaagtc acttttgaca 2461 ttgatgccaa tggtatactc aatgtctctg ctgtggacaa gagtacggga aaagagaaca 2521 agattactat cactaatgac aagggccgtt tgagcaagga agacattgaa cgtatggtcc 2581 aggaagctga gaagtacaaa gctgaagatg agaagcagag ggacaaggtg tcatccaaga 2641 attcacttga gtcctatgcc ttcaacatga aagcaactgt tgaagatgag aaacttcaag 2701 gcaagattaa cgatgaggac aaacagaaga ttctggacaa gtgtaatgaa attatcaact 2761 ggcttgataa gaatcagact gctgagaagg aagaatttga acatcaacag aaagagctgg 2821 agaaagtttg caaccccatc atcaccaagc tgtaccagag tgcaggaggc atgccaggag 2881 gaatgcctgg gggatttcct ggtggtggag ctcctccctc tggtggtgct tcctcagggc 2941 ccaccattga agaggttgat taagcggccg ctcgagtcta gagggcccgt ttaaacccgc 3001 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 3061 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 3121 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 3181 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 3241 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 3301 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3361 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3421 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 3481 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt 3541 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg 3601 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg 3661 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg 3721 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg 3781 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat 3841 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc 3901 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag 3961 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat 4021 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc 4081 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc 4141 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc 4201 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc 4261 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc 4321 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt 4381 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg 4441 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc 4501 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga 4561 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat 4621 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 4681 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 4741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 4801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 4861 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 4921 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 4981 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 5041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 5101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 5161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 5221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 5281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 5341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 5401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 5461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 5521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 5581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 5641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 5701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 5761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 5821 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 5881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 5941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 6001 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 6061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 6121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 6181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 6241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 6301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 6361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 6421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 6481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 6541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 6601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 6661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 6721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 6781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 6841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 6901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 6961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 7021 ccgcgcacat ttccccgaaa agtgccacct gacgtc //