LOCUS Exported 6291 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO DNAJA1 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 12, 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" enhancer 123..502 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 503..706 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 657..677 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 708..726 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 729..747 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 757..781 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind complement(2131..2148) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2137..2361 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2494..2513) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2645..2692 /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 2700..3722 /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 3852..3973 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3889..3908) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3943..3962 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4022..4038) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4022..4038) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4035..4057) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4046..4062 /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(4070..4100) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4115..4136 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4253..4270) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4424..5012) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4504..4523) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5183..6043) /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 5806..5825 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6044..6148) /gene="bla" /label=AmpR promoter primer_bind complement(6223..6242) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa 61 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg 121 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag 181 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc 241 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg 301 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca 361 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc 421 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt 481 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata 541 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt 601 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca 661 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga 721 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat 781 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag 841 cctccggact ctagcgttta aacttaagct tggtaccgag ctcggatcca ccatggtgaa 901 agaaacaact tactacgatg ttttgggggt caaacccaat gctactcagg aagaattgaa 961 aaaggcttat aggaaactgg ctttgaagta ccatcctgat aagaacccaa atgaaggaga 1021 gaagtttaaa cagatttctc aagcttacga agttctctct gatgcaaaga aaagggaatt 1081 atatgacaaa ggaggagaac aggcaattaa agagggtgga gcaggtggcg gttttggctc 1141 ccccatggac atctttgata tgttttttgg aggaggagga aggatgcaga gagaaaggag 1201 aggtaaaaat gttgtacatc agctctcagt aaccctagaa gacttatata atggtgcaac 1261 aagaaaactg gctctgcaaa agaatgtgat ttgtgacaaa tgtgaaggta gaggaggtaa 1321 gaaaggagca gtagagtgct gtcccaattg ccgaggtact ggaatgcaaa taagaattca 1381 tcagatagga cctggaatgg ttcagcaaat tcagtctgtg tgcatggagt gccagggcca 1441 tggggagcgg atcagtccta aagatagatg taaaagctgc aacggaagga agatagttcg 1501 agagaagaaa attttagaag ttcatattga caaaggcatg aaagatggcc agaagataac 1561 attccatggt gaaggagacc aagaaccagg actggagcca ggcgatatta tcattgtgtt 1621 aggtcagaag gaccatgctg tttttactcg acgaggagaa gaccttttca tgtgtatgga 1681 catacagctc gttgaagcac tgtgtggctt ccagaagcca atatctactc ttgacaaccg 1741 aaccatcgtc atcacctctc atccaggtca gattgtcaag catggagata tcaagtgtgt 1801 actaaatgaa ggcatgccaa tttatcgtag accatatgaa aagggtcgcc taatcatcga 1861 atttaaggta aactttcctg agaatggctt tctctctcct gataaactgt ctttgctgga 1921 aaaactccta cccgagagga aggaagtgga agagactgat gagatggacc aagtagaact 1981 ggtggacttt gatccaaatc aggaaagacg gcgccactac aatggagaag catatgagga 2041 tgatgaacat catcccagag gtggtgttca gtgtcagacc tcttaagcgg ccgctcgagt 2101 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat 2161 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc 2221 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg 2281 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg 2341 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt 2401 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 2461 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 2521 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 2581 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 2641 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca 2701 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg 2761 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag 2821 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt 2881 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg 2941 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag 3001 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga 3061 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg 3121 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact 3181 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga 3241 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca 3301 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt 3361 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta 3421 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc 3481 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca 3541 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg 3601 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg 3661 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat 3721 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 3781 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 3841 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 3901 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 3961 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 4021 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4081 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4141 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4201 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4261 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4321 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4381 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4441 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4501 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4561 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 4621 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 4681 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 4741 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 4801 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 4861 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 4921 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 4981 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 5041 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 5101 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 5161 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 5221 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 5281 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 5341 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 5401 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 5461 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 5521 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 5581 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 5641 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 5701 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 5761 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 5821 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 5881 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 5941 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 6001 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 6061 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 6121 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 6181 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc 6241 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg a //