LOCUS Exported 7098 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO V5 HSPA1L SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7098) 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 7098) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7098 /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(3050..3067) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3056..3280 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3413..3432) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3564..3611 /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 3619..4641 /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 4771..4892 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4808..4827) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4862..4881 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4941..4957) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4941..4957) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4954..4976) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4965..4981 /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(4989..5019) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5034..5055 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5172..5189) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5343..5931) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5423..5442) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6102..6962) /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 6725..6744 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6963..7067) /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 tccactagtc cagtgtggtg gaattctgca gatatcacca 1081 tggctactgc caagggaatc gccataggca tcgacctggg caccacctac tcctgtgtgg 1141 gggtgttcca gcacggcaag gtggagatca tcgccaacga ccagggcaac cgcaccaccc 1201 ccagctacgt ggccttcaca gacaccgagc ggctcattgg ggatgcggcc aagaaccagg 1261 tagcaatgaa tccccagaac actgtttttg atgctaaacg tctgatcggc aggaaattta 1321 atgatcctgt tgtacaagca gatatgaaac tttggccttt tcaagtgatt aatgaaggag 1381 gcaagcccaa agtccttgtg tcctacaaag gggagaataa agctttctac cctgaggaaa 1441 tctcttcgat ggtattgact aagttgaagg agactgctga ggcctttttg ggccaccctg 1501 tcaccaatgc agtgattacc gtgccagcct atttcaatga ctctcaacgt caggctacta 1561 aggatgcagg tgtgattgct ggacttaatg tgctaagaat catcaatgag cccacggctg 1621 ctgccattgc ctatggttta gataaaggag gtcaaggaga acgacatgtc ctgatttttg 1681 atctgggtgg aggcacattt gatgtgtcaa ttctgaccat agatgatggg atttttgagg 1741 taaaggccac tgctggggac actcacctgg gtggggagga ctttgacaac aggcttgtga 1801 gccacttcgt ggaggagttc aagaggaaac acaaaaagga catcagccag aacaagcgag 1861 ccgtgaggcg gctgcgcacc gcctgcgaga gggccaagag gaccctgtcg tccagcaccc 1921 aggccaacct agaaattgat tcactttatg aaggcattga cttctataca tccatcacca 1981 gagctcgatt tgaagagttg tgtgcagacc tgtttagggg taccctggag cctgtagaaa 2041 aagcgcttcg ggatgccaag atggataagg ctaaaatcca tgacattgtt ttagtagggg 2101 gctccacccg catccccaag gtgcagcggc tgcttcagga ctacttcaat ggacgtgatc 2161 tcaacaagag catcaaccct gatgaggccg tagcatatgg ggctgcggta caagcagcca 2221 tcctgatggg ggacaagtct gagaaggtac aggacctgct gctgctggac gtggctcccc 2281 tgtccctggg gctggagacg gctgggggcg tgatgactgc cctgataaag cgcaactcca 2341 ccatccccac caagcagaca cagattttca ccacctactc tgacaaccaa cccggggtgc 2401 tgatccaggt gtatgagggc gagagggcca tgacaaagga caacaacctg ctggggcggt 2461 ttgacctgac tggaatccct ccagcaccca ggggagttcc tcagatcgag gtgacgtttg 2521 acattgatgc caatggtatt ctcaatgtca cagccacgga caagagcacc ggcaaggtga 2581 acaagatcac catcaccaat gacaagggcc gcctgagcaa ggaggagatt gagcgcatgg 2641 ttctggatgc tgagaaatat aaagctgaag atgaggtcca gagggagaaa attgctgcaa 2701 agaatgcctt agaatcctat gcttttaaca tgaagagtgt tgtgagtgat gaaggtttga 2761 agggcaagat tagtgagtct gataaaaata aaatattgga taaatgcaac gagctccttt 2821 cgtggctgga ggtcaatcaa ctggcagaga aagatgagtt tgatcataag agaaaggaat 2881 tggagcagat gtgtaaccct atcatcacaa aactctacca aggaggatgc actgggcctg 2941 cctgcggaac agggtatgtg cctggaaggc ctgccacagg ccccacaatt gaagaagtag 3001 attaagcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg 3061 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 3121 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 3181 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 3241 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 3301 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3541 tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt 3601 ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg 3661 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt 3721 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat 3781 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg 3841 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca 3901 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc 3961 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca 4021 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct 4081 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg 4141 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg 4201 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt 4261 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg 4321 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag 4381 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat 4441 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca 4501 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc 4561 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact 4621 cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat 4681 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 4741 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 4801 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 4861 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 4921 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 4981 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 5041 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 5101 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 5161 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 5221 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 5281 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 5341 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 5401 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 5461 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 5521 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 5581 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 5641 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 5701 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 5761 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 5821 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 5881 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 5941 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 6001 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 6061 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 6121 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 6181 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 6241 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 6301 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 6361 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 6421 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 6481 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 6541 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 6601 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 6661 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 6721 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 6781 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 6841 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 6901 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 6961 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 7021 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 7081 aaagtgccac ctgacgtc //