LOCUS Exported 7056 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO HSPA1L 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" 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 cttggtaccg agctcggatc cactagtcca gtgtggtgga 1021 attctgcaga tatcaccatg gctactgcca agggaatcgc cataggcatc gacctgggca 1081 ccacctactc ctgtgtgggg gtgttccagc acggcaaggt ggagatcatc gccaacgacc 1141 agggcaaccg caccaccccc agctacgtgg ccttcacaga caccgagcgg ctcattgggg 1201 atgcggccaa gaaccaggta gcaatgaatc cccagaacac tgtttttgat gctaaacgtc 1261 tgatcggcag gaaatttaat gatcctgttg tacaagcaga tatgaaactt tggccttttc 1321 aagtgattaa tgaaggaggc aagcccaaag tccttgtgtc ctacaaaggg gagaataaag 1381 ctttctaccc tgaggaaatc tcttcgatgg tattgactaa gttgaaggag actgctgagg 1441 cctttttggg ccaccctgtc accaatgcag tgattaccgt gccagcctat ttcaatgact 1501 ctcaacgtca ggctactaag gatgcaggtg tgattgctgg acttaatgtg ctaagaatca 1561 tcaatgagcc cacggctgct gccattgcct atggtttaga taaaggaggt caaggagaac 1621 gacatgtcct gatttttgat ctgggtggag gcacatttga tgtgtcaatt ctgaccatag 1681 atgatgggat ttttgaggta aaggccactg ctggggacac tcacctgggt ggggaggact 1741 ttgacaacag gcttgtgagc cacttcgtgg aggagttcaa gaggaaacac aaaaaggaca 1801 tcagccagaa caagcgagcc gtgaggcggc tgcgcaccgc ctgcgagagg gccaagagga 1861 ccctgtcgtc cagcacccag gccaacctag aaattgattc actttatgaa ggcattgact 1921 tctatacatc catcaccaga gctcgatttg aagagttgtg tgcagacctg tttaggggta 1981 ccctggagcc tgtagaaaaa gcgcttcggg atgccaagat ggataaggct aaaatccatg 2041 acattgtttt agtagggggc tccacccgca tccccaaggt gcagcggctg cttcaggact 2101 acttcaatgg acgtgatctc aacaagagca tcaaccctga tgaggccgta gcatatgggg 2161 ctgcggtaca agcagccatc ctgatggggg acaagtctga gaaggtacag gacctgctgc 2221 tgctggacgt ggctcccctg tccctggggc tggagacggc tgggggcgtg atgactgccc 2281 tgataaagcg caactccacc atccccacca agcagacaca gattttcacc acctactctg 2341 acaaccaacc cggggtgctg atccaggtgt atgagggcga gagggccatg acaaaggaca 2401 acaacctgct ggggcggttt gacctgactg gaatccctcc agcacccagg ggagttcctc 2461 agatcgaggt gacgtttgac attgatgcca atggtattct caatgtcaca gccatggaca 2521 agagcaccgg caaggtgaac aagatcacca tcaccaatga caagggccgc ctgagcaagg 2581 aggagattga gcgcatggtt ctggatgctg agaaatataa agctgaagat gaggtccaga 2641 gggagaaaat tgctgcaaag aatgccttag aatcctatgc ttttaacatg aagagtgttg 2701 tgagtgatga aggtttgaag ggcaagatta gtgagtctga taaaaataaa atattggata 2761 aatgcaacga gctcctttcg tggctggagg tcaatcaact ggcagagaaa gatgagtttg 2821 atcataagag aaaggaattg gagcagatgt gtaaccctat catcacaaaa ctctaccaag 2881 gaggatgcac tgggcctgcc tgcggaacag ggtatgtgcc tggaaggcct gccacaggcc 2941 ccacaattga agaagtagat 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 //