LOCUS Exported 6645 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO HIS HSPA14 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6645) 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 6645) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6645 /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" regulatory 1021..1030 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(2597..2614) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2603..2827 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2960..2979) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3111..3158 /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 3166..4188 /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 4318..4439 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4355..4374) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4409..4428 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4488..4504) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4488..4504) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4501..4523) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4512..4528 /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(4536..4566) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4581..4602 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4719..4736) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4890..5478) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4970..4989) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5649..6509) /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 6272..6291 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6510..6614) /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 ccatggcggc catcggagtt cacctgggct gcacctcagc ctgtgtggcc gtctataagg 1081 atggccgggc tggtgtggtt gcaaatgatg ccggtgaccg agttactcca gctgttgttg 1141 cttactcaga aaatgaagag attgttggat tggcagcaaa acaaagtaga ataagaaata 1201 tttcaaatac agtaatgaaa gtaaagcaga tcctgggcag aagctccagt gatccacaag 1261 ctcagaaata catcgcggaa agtaaatgtt tagtcattga aaaaaatggg aaattacgat 1321 atgaaataga tactggagaa gaaacaaaat ttgttaaccc agaagatgtt gccagactga 1381 tatttagtaa aatgaaagaa acggcacatt ctgtattggg ctcagatgca aatgatgtag 1441 ttattactgt cccgtttgat tttggagaaa agcaaaaaaa tgcccttgga gaagcagcta 1501 gagctgctgg atttaatgtt ttgcgattaa ttcacgaacc gtctgcagct cttcttgctt 1561 atggaattgg acaagactcc cctactggaa aaagcaatat tttggtgttt aagcttggag 1621 gaacatcctt atctctcagc gtcatggaag ttaacagtgg aatatatcgg gttctttcaa 1681 caaacactga tgataacatc ggtggtgcac atttcacaga aaccttagca cagtatctag 1741 cttctgagtt ccaaagatcc ttcaaacatg atgtgagagg aaatgcgcga gccatgatga 1801 aattaacgaa cagtgctgaa gtagcgaaac attctttgtc aaccttggga agtgccaact 1861 gttttcctga ctcattatat gaagggcaag attttgattg caatgtgtcc agagcaagat 1921 ttgaacttct ttgttctcca ctttttaata agtgtttaga agcaatcaga ggactcttag 1981 atcaaaatgg atttacagca gatgatatca acaaggttgt cctttgtgga gggtcttctc 2041 gaatcccaaa gctacagcaa ctgattaaag atcttttccc agctgttgag cttctcaatt 2101 ctatccctcc tgatgaagtg atccctattg gtgcagctat agaagcagga attcttattg 2161 ggaaagaaaa cctgttggtg gaagactctc ttatgataga gtgttcagcc agagatattt 2221 tagttaaggg tgtggacgaa tcaggagcca gtagattcac agtgctgttt ccatcaggga 2281 ctcctttgcc agctcgaaga caacacacat tgcaagcccc tggaagcata tcttcagtgt 2341 gccttgaact ctatgagtct gatgggaaga actctgccaa agaggaaacc aagtttgcac 2401 aggttgtact ccaggattta gataaaaaag aaaatggatt acgtgatata ttagctgttc 2461 ttactatgaa aagggatgga tctttacatg tgacatgcac agatcaagaa actggaaaat 2521 gtgaagcaat ctctattgag atagcatctt aggcggccgc tcgagtctag agggcccgtt 2581 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc 2641 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat 2701 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg 2761 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc 2821 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc 2881 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 2941 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 3001 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 3061 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta gaagttccta 3121 ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa gcctgaactc 3181 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg 3241 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat 3301 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac 3361 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc 3421 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc 3481 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat 3541 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca 3601 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg 3661 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag 3721 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg 3781 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa 3841 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg 3901 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg 3961 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca 4021 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt 4081 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc 4141 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga 4201 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 4261 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 4321 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 4381 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 4441 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 4501 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 4561 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 4621 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 4681 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 4741 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 4801 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 4861 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4921 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4981 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 5041 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 5101 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 5161 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 5221 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 5281 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 5341 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 5401 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 5461 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 5521 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 5581 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 5641 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 5701 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 5761 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 5821 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5881 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5941 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6001 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 6061 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 6121 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6181 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6241 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6301 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 6361 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 6421 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 6481 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 6541 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 6601 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //