LOCUS Exported 7041 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO HIS HSPA1A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7041) 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 7041) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7041 /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(2993..3010) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2999..3223 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3356..3375) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3507..3554 /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 3562..4584 /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 4714..4835 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4751..4770) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4805..4824 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4884..4900) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4884..4900) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4897..4919) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4908..4924 /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(4932..4962) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4977..4998 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5115..5132) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5286..5874) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5366..5385) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6045..6905) /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 6668..6687 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6906..7010) /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 ccatggccaa agccgcggcg atcggcatcg acctgggcac cacctactcc tgcgtggggg 1081 tgttccaaca cggcaaggtg gagatcatcg ccaacgacca gggcaaccgc accaccccca 1141 gctacgtggc cttcacggac accgagcggc tcatcgggga tgcggccaag aaccaggtgg 1201 cgctgaaccc gcagaacacc gtgtttgacg cgaagcggct gatcggccgc aagttcggcg 1261 acccggtggt gcagtcggac atgaagcact ggcctttcca ggtgatcaac gacggagaca 1321 agcccaaggt gcaggtgagc tacaaggggg acaccaaggc attctacccc gaggagatct 1381 cgtccatggt gctgaccaag atgaaggaga tcgccgaggc gtacctgggc tacccggtga 1441 ccaacgcggt gatcaccgtg ccggcctact tcaacgactc gcagcgccag gccaccaagg 1501 atgcgggtgt gatcgcgggg ctcaacgtgc tgcggatcat caacgagccc acggccgccg 1561 ccatcgccta cggcctggac agaacgggca agggggagcg caacgtgctc atctttgacc 1621 tgggcggggg caccttcgac gtgtccatcc tgacgatcga cgacggcatc ttcgaggtga 1681 aggccacggc cggggacacc cacctgggtg gggaggactt tgacaacagg ctggtgaacc 1741 acttcgtgga ggagttcaag agaaaacaca agaaggacat cagccagaac aagcgagccg 1801 tgaggcggct gcgcaccgcc tgcgagaggg ccaagaggac cctgtcgtcc agcacccagg 1861 ccagcctgga gatcgactcc ctgtttgagg gcatcgactt ctacacgtcc atcaccaggg 1921 cgaggttcga ggagctgtgc tccgacctgt tccgaagcac cctggagccc gtggagaagg 1981 ctctgcgcga cgccaagctg gacaaggccc agattcacga cctggtcctg gtcgggggct 2041 ccacccgcat ccccaaggtg cagaagctgc tgcaggactt cttcaacggg cgcgacctga 2101 acaagagcat caaccccgac gaggctgtgg cctacggggc ggcggtgcag gcggccatcc 2161 tgatggggga caagtccgag aacgtgcagg acctgctgct gctggacgtg gctcccctgt 2221 cgctggggct ggagacggcc ggaggcgtga tgactgccct gatcaagcgc aactccacca 2281 tccccaccaa gcagacgcag atcttcacca cctactccga caaccaaccc ggggtgctga 2341 tccaggtgta cgagggcgag agggccatga cgaaagacaa caatctgttg gggcgcttcg 2401 agctgagcgg catccctccg gcccccaggg gcgtgcccca gatcgaggtg accttcgaca 2461 tcgatgccaa cggcatcctg aacgtcacgg ccacggacaa gagcaccggc aaggccaaca 2521 agatcaccat caccaacgac aagggccgcc tgagcaagga ggagatcgag cgcatggtgc 2581 aggaggcgga gaagtacaaa gcggaggacg aggtgcagcg cgagagggtg tcagccaaga 2641 acgccctgga gtcctacgcc ttcaacatga agagcgccgt ggaggatgag gggctcaagg 2701 gcaagatcag cgaggccgac aagaagaagg tgctggacaa gtgtcaagag gtcatctcgt 2761 ggctggacgc caacaccttg gccgagaagg acgagtttga gcacaagagg aaggagctgg 2821 agcaggtgtg taaccccatc atcagcggac tgtaccaggg tgccggtggt cccgggcctg 2881 ggggcttcgg ggctcagggt cccaagggag ggtctgggtc aggccccacc attgaggagg 2941 tagattaggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact 3001 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 3061 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 3121 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 3181 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 3241 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 3301 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 3361 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 3421 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 3481 gattagggtg atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa 3541 agtataggaa cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt 3601 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct 3661 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc 3721 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt 3781 ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt 3841 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg 3901 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc 3961 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt 4021 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc 4081 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc 4141 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc 4201 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc 4261 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg 4321 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc 4381 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac 4441 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg 4501 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc 4561 actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc 4621 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 4681 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 4741 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 4801 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 4861 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 4921 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 4981 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 5041 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 5101 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 5161 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 5221 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 5281 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 5341 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 5401 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 5461 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 5521 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 5581 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 5641 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 5701 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 5761 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 5821 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 5881 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 5941 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 6001 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 6061 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 6121 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 6181 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 6241 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 6301 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 6361 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 6421 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 6481 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 6541 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 6601 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 6661 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 6721 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 6781 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 6841 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 6901 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 6961 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 7021 cgaaaagtgc cacctgacgt c //