LOCUS Exported 6822 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO GFP DNAJB4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6822) 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 6822) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6822 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 6..15 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 12..728 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 12..728 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(57..78) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(318..337) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 665..686 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1796..1813) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1802..2026 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2159..2178) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2310..2357 /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 2365..3387 /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 3517..3638 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3554..3573) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3608..3627 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3687..3703) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3687..3703) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3700..3722) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3711..3727 /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(3735..3765) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3780..3801 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3918..3935) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4089..4677) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4169..4188) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4848..5708) /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 5471..5490 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5709..5813) /gene="bla" /label=AmpR promoter primer_bind complement(5888..5907) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 6079..6458 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6459..6662 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6613..6633 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 6664..6682 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 6685..6703 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 6713..6737 /label=LNCX /note="Human CMV promoter, forward primer" ORIGIN 1 agcttgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg 61 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 121 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc 181 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg 241 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 301 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 361 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 421 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca 481 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg 541 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 601 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg 661 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc 721 tgtacaaggg atccaccatg gggaaagact attattgcat tttgggaatt gagaaaggag 781 cttcagatga agatattaaa aaggcttacc gaaaacaagc cctcaaattt catccggaca 841 agaacaaatc tcctcaggca gaggaaaaat ttaaagaggt cgcagaagct tatgaagtat 901 tgagtgatcc taaaaagaga gaaatatatg atcagtttgg ggaggaaggg ttgaaaggag 961 gagcaggagg tactgatgga caaggaggta ccttccggta cacctttcat ggcgatcctc 1021 atgctacatt tgctgcattt ttcggagggt ccaacccctt tgaaattttc tttggaagac 1081 gaatgggtgg tggtagagat tctgaagaaa tggaaataga tggtgatcct tttagtgcct 1141 ttggtttcag catgaatgga tatccaagag acaggaattc tgtggggcca tcccgcctca 1201 aacaagatcc tccagttatt catgaactta gagtatcact tgaagagata tatagtggtt 1261 gtaccaaacg gatgaagatt tctcgaaaaa ggctaaacgc tgatggaagg agttacagat 1321 ctgaggacaa aattcttacc attgagatta aaaaagggtg gaaagaaggc accaaaatta 1381 cttttccaag agaaggagat gaaacaccaa atagtattcc agcagacatt gtttttatca 1441 ttaaagacaa agatcatcca aaatttaaaa gggatggatc aaatataatt tatactgcta 1501 aaattagttt acgagaggca ttgtgtggct gctcaattaa tgtaccaaca ctggatggaa 1561 gaaacatacc tatgtcagta aatgatattg tgaaacccgg aatgaggaga agaattattg 1621 gatatgggct gccatttcca aaaaatcctg accaacgtgg tgaccttcta atagaatttg 1681 aggtgtcctt cccagatact atatcttctt catccaaaga agtacttagg aaacatcttc 1741 ctgcctcata ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg 1801 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 1861 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 1921 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 1981 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 2041 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 2101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 2221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 2281 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta 2341 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag 2401 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa 2461 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc 2521 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg 2581 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc 2641 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag 2701 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc 2761 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg 2821 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc 2881 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac 2941 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg 3001 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc 3061 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat 3121 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa 3181 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc 3241 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc 3301 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc 3361 agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc 3421 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 3481 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 3541 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 3601 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 3661 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 3721 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 3781 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 3841 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 3901 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 3961 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 4021 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 4081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 4141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 4201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 4261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 4321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 4381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 4441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 4501 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 4561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 4621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 4681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 4741 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 4801 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 4861 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 4921 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 4981 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 5041 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 5101 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 5161 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 5221 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 5281 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 5341 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 5401 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 5461 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 5521 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 5581 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 5641 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 5701 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 5761 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 5821 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca 5881 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg 5941 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga 6001 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac 6061 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg 6121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc 6181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca 6241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 6301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg 6361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt 6421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca 6481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 6541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact 6601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 6661 ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga cgagctcgtt 6721 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac 6781 accgggaccg atccagcctc cggactctag cgtttaaact ta //