LOCUS Exported 6291 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO DNAJA4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6291) 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 6291) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6291 /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(2243..2260) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2249..2473 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2606..2625) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2757..2804 /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 2812..3834 /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 3964..4085 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4001..4020) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4055..4074 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4134..4150) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4134..4150) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4147..4169) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4158..4174 /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(4182..4212) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4227..4248 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4365..4382) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4536..5124) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4616..4635) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5295..6155) /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 5918..5937 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6156..6260) /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 caccatggtg aaggagaccc 1021 agtactatga catcctgggc gtgaagccca gcgcgtcccc ggaggagatc aagaaggcct 1081 atcggaagct ggcgctcaag tgccacccgg acaagaaccc ggatgagggc gagaagttta 1141 aactcatatc ccaggcatat gaagtgcttt cagatccaaa gaaaagggat gtttatgacc 1201 aaggcggaga gcaggcaatt aaagaaggag gctcaggcag ccccagcttc tcttcaccca 1261 tggacatctt tgacatgttc tttggtggtg gtggacggat ggctagagag agaagaggca 1321 agaatgttgt acaccagtta tctgtaactc ttgaagattt atataatgga gtcacgaaga 1381 aattggccct ccagaaaaat gtaatttgtg agaaatgtga aggtgttggt gggaagaagg 1441 gatcggtgga gaagtgcccg ctgtgcaagg ggcgggggat gcagatccac atccagcaga 1501 tcgggccggg catggtacag cagatccaga ccgtgtgcat cgagtgcaag ggccagggtg 1561 agcgcatcaa ccccaaggac cgctgcgaga gctgcagcgg ggccaaggtg atccgtgaga 1621 agaagattat cgaggtacat gttgaaaaag gtatgaaaga tgggcaaaag atactatttc 1681 atggagaagg agatcaggag cctgagctgg agcctggtga tgtcataatt gtgcttgatc 1741 agaaggatca tagtgtcttt cagagacgag gccatgactt gatcatgaaa atgaaaattc 1801 agctttctga agctctttgt ggcttcaaga agacgataaa aacattggac aatcgaattc 1861 ttgttattac atccaaagca ggtgaggtga taaagcacgg ggacctgaga tgcgtgcgcg 1921 atgaaggaat gcccatctac aaagcacccc tggaaaaagg gattctgatc atacagtttt 1981 tagtaatctt tcctgaaaaa cactggcttt ctctggaaaa gcttcctcag ctggaagctt 2041 tactccctcc tcgacagaaa gtgaggatta cagatgacat ggatcaggtg gagctgaagg 2101 agttttgtcc caatgagcag aactggcgtc agcacaggga ggcctacgag gaggacgaag 2161 acgggcccca ggctggagtg cagtgccaga cggcatgagc ggccgctcga gtctagaggg 2221 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 2281 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 2341 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 2401 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 2461 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac 2521 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 2581 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 2641 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 2701 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tacctagaag 2761 ttcctattcc gaagttccta ttctctagaa agtataggaa cttccttggc caaaaagcct 2821 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac 2881 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt 2941 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat 3001 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc 3061 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct 3121 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg 3181 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac 3241 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact 3301 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg 3361 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc 3421 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat 3481 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag 3541 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg 3601 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat 3661 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc 3721 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta 3781 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta 3841 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 3901 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 3961 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 4021 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 4081 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 4141 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 4201 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 4261 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 4321 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 4381 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 4441 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 4501 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4561 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4621 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4681 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 4741 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4801 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 4861 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 4921 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 4981 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5041 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5101 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5161 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 5221 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 5281 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5341 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5401 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 5461 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 5521 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 5581 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5641 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5701 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5761 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 5821 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 5881 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 5941 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6001 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6061 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6121 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6181 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 6241 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //