LOCUS Exported 7059 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO V5 HSPA2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7059) 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 7059) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7059 /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..1037 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3011..3028) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3017..3241 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3374..3393) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3525..3572 /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 3580..4602 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVT*QDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4732..4853 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4769..4788) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4823..4842 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4902..4918) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4902..4918) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4915..4937) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4926..4942 /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(4950..4980) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4995..5016 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5133..5150) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5304..5892) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5384..5403) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6063..6923) /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 6686..6705 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6924..7028) /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 cttgaattca ccatgggtaa gcctatccct aaccctctcc 1021 tcggtctcga ttctacggga tccaccatgt ctgcccgtgg cccggctatc ggcatcgacc 1081 tgggcaccac ctattcgtgc gtcggggtct tccaacatgg caaggtggag atcatcgcca 1141 acgaccaggg caatcgcacc acccccagct acgtggcctt cacggacacc gagcgcctca 1201 tcggcgacgc cgccaagaac caggtggcca tgaaccccac caacaccatc ttcgacgcca 1261 agaggctgat tggacggaaa ttcgaggatg ccacagtgca gtcggatatg aaacactggc 1321 cgttccgggt ggtgagcgag ggaggcaagc ccaaagtgca agtagagtac aagggggaga 1381 ccaagacctt cttcccagag gagatatcct ccatggtcct cacgaagatg aaggagatcg 1441 cggaagccta cctggggggc aaggtgcaca gcgcggtcat aacggtcccg gcctatttca 1501 acgactcgca gcgccaggcc accaaggacg caggcaccat cacggggctc aatgtgctgc 1561 gcatcatcaa cgagcccacg gcggcggcca tcgcctacgg cctggacaag aagggctgcg 1621 cgggcggcga gaagaacgtg ctcatctttg acctgggcgg tggcactttc gacgtgtcca 1681 tcctgaccat cgaggatggc atcttcgagg tgaagtccac ggccggcgat acccacctgg 1741 gcggtgagga cttcgacaac cgcatggtga gccacctggc ggaggagttc aagcgcaagc 1801 acaagaagga cattgggccc aacaagcgcg ccgtgaggcg gctgcgcacc gcttgcgagc 1861 gcgccaagcg caccctgagc tcgtccacgc aggcgagcat cgagatcgac tcgctctacg 1921 agggcgtgga cttctatacg tccatcacgc gcgcccgctt cgaggagctc aatgccgacc 1981 tctttcgcgg gaccctggag ccggtggaga aggcgctgcg cgacgccaag ctggacaagg 2041 gccagatcca ggagatcgtg ctggtgggcg gctccactcg tatccccaag atccagaagc 2101 tgctgcagga tttcttcaac ggcaaggagc tgaacaagag catcaacccc gacgaggcgg 2161 tggcctatgg cgccgcggtg caggcggcca tcctcatcgg cgacaaatca gagaatgtgc 2221 aggacctgct gctactcgac gtgaccccgt tgtcgctggg catcgagaca gctggcggtg 2281 tcatgacccc actcatcaag aggaacacca cgatccccac caagcagacg cagaccttca 2341 ccacctactc ggacaaccag agcagcgtac tggtgcaggt atacgagggc gaacgggcca 2401 tgaccaagga caataacctg ctgggcaagt tcgacctgac cgggattccc cctgcgcctc 2461 gcggggtccc ccaaatcgag gttaccttcg acattgacgc caatggcatc cttaacgtta 2521 ccgccgccga caagagcacc ggtaaggaaa acaaaatcac catcaccaat gacaaaggtc 2581 gtctgagcaa ggacgacatt gaccggatgg tgcaggaggc ggagcggtac aaatcggaag 2641 atgaggcgaa tcgcgaccga gtcgcggcca aaaacgccct ggagtcctat acctacaaca 2701 tcaagcagac ggtggaagac gagaaactga ggggcaagat tagcgagcag gacaaaaaca 2761 agatcctcga caagtgtcag gaggtgatca actggctcga ccgaaaccag atggcagaga 2821 aagatgagta tgaacacaag cagaaagagc tcgaaagagt ttgcaacccc atcatcagca 2881 aactttacca aggtggtcct ggcggcggca gcggcggcgg cggttcagga gcctccgggg 2941 gacccaccat cgaagaagtg gactaagcgg ccgctcgagt ctagagggcc cgtttaaacc 3001 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 3061 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 3121 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 3181 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 3241 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 3301 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 3361 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 3421 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 3481 ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga 3541 agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg 3601 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc 3661 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg 3721 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca 3781 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc 3841 tattgcatct cccgccgtgc acagggtgtc acgtagcaag acctgcctga aaccgaactg 3901 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc 3961 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt 4021 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac 4081 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc 4141 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat 4201 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag 4261 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac 4321 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc 4381 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg 4441 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa 4501 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt 4561 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc 4621 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 4681 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 4741 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4801 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 4861 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 4921 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 4981 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5041 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5101 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5161 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5401 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5461 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 5521 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5581 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5641 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5701 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 5761 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5821 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 5881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 5941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6061 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6121 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6181 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6241 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6301 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6361 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6421 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6481 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6541 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6601 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6661 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6721 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6781 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6841 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6901 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 6961 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7021 gttccgcgca catttccccg aaaagtgcca cctgacgtc //