LOCUS Exported 7728 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA5/FRT/TO GFP HSPA2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7728) 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 7728) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7728 /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(2702..2719) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2708..2932 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3065..3084) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3216..3263 /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 3271..4293 /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 4423..4544 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4460..4479) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4514..4533 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4593..4609) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4593..4609) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4606..4628) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4617..4633 /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(4641..4671) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4686..4707 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4824..4841) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4995..5583) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5075..5094) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5754..6614) /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 6377..6396 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6615..6719) /gene="bla" /label=AmpR promoter primer_bind complement(6794..6813) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 6985..7364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7365..7568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7519..7539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 7570..7588 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 7591..7609 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 7619..7643 /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 tctgcccgtg gcccggctat cggcatcgac ctgggcacca 781 cctattcgtg cgtcggggtc ttccaacatg gcaaggtgga gatcatcgcc aacgaccagg 841 gcaatcgcac cacccccagc tacgtggcct tcacggacac cgagcgcctc atcggcgacg 901 ccgccaagaa ccaggtggcc atgaacccca ccaacaccat cttcgacgcc aagaggctga 961 ttggacggaa attcgaggat gccacagtgc agtcggatat gaaacactgg ccgttccggg 1021 tggtgagcga gggaggcaag cccaaagtgc aagtagagta caagggggag accaagacct 1081 tcttcccaga ggagatatcc tccatggtcc tcacgaagat gaaggagatc gcggaagcct 1141 acctgggggg caaggtgcac agcgcggtca taacggtccc ggcctatttc aacgactcgc 1201 agcgccaggc caccaaggac gcaggcacca tcacggggct caatgtgctg cgcatcatca 1261 acgagcccac ggcggcggcc atcgcctacg gcctggacaa gaagggctgc gcgggcggcg 1321 agaagaacgt gctcatcttt gacctgggcg gtggcacttt cgacgtgtcc atcctgacca 1381 tcgaggatgg catcttcgag gtgaagtcca cggccggcga cacccacctg ggcggtgagg 1441 acttcgacaa ccgcatggtg agccacctgg cggaggagtt caagcgcaag cacaagaagg 1501 acattgggcc caacaagcgc gccgtgaggc ggctgcgcac cgcttgcgag cgcgccaagc 1561 gcaccctgag ctcgtccacg caggcgagca tcgagatcga ctcgctctac gagggcgtgg 1621 acttctatac gtccatcacg cgcgcccgct tcgaggagct caatgccgac ctctttcgcg 1681 ggaccctgga gccggtggag aaggcgctgc gcgacgccaa gctggacaag ggccagatcc 1741 aggagatcgt gctggtgggc ggctccactc gtatccccaa gatccagaag ctgctgcagg 1801 atttcttcaa cggcaaggag ctgaacaaga gcatcaaccc cgacgaggcg gtggcctatg 1861 gcgccgcggt gcaggcggcc atcctcatcg gcgacaaatc agagaatgtg caggacctgc 1921 tgctactcga cgtgaccccg ttgtcgctgg gcatcgagac agctggcggt gtcatgaccc 1981 cactcatcaa gaggaacacc acgatcccca ccaagcagac gcagaccttc accacctact 2041 cggacaacca gagcagcgta ctggtgcagg tatacgaggg cgaacgggcc atgaccaagg 2101 acaataacct gctgggcaag ttcgacctga ccgggattcc ccctgcgcct cgcggggtcc 2161 cccaaatcga ggttaccttc gacattgacg ccaatggcat ccttaacgtt accgccgccg 2221 acaagagcac cggtaaggaa aacaaaatca ccatcaccaa tgacaaaggt cgtctgagca 2281 aggacgacat tgaccggatg gtgcaggagg cggagcggta caaatcggaa gatgaggcga 2341 atcgcgaccg agtcgcggcc aaaaacgccc tggagtccta tacctacaac atcaagcaga 2401 cggtggaaga cgagaaactg aggggcaaga ttagcgagca ggacaaaaac aagatcctcg 2461 acaagtgtca ggaggtgatc aactggctcg accgaaacca gatggcagag aaagatgagt 2521 atgaacacaa gcagaaagag ctcgaaagag tttgcaaccc catcatcagc aaactttacc 2581 aaggtggtcc tggcggcggc agcggcggcg gcggttcagg agcctccggg ggacccacca 2641 tcgaagaagt ggactaagcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca 2701 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 2761 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 2821 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 2881 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 2941 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 3001 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 3061 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 3121 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 3181 aaaaaacttg attagggtga tggttcacgt acctagaagt tcctattccg aagttcctat 3241 tctctagaaa gtataggaac ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc 3301 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc 3361 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat 3421 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg 3481 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc 3541 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt 3601 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc 3661 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata 3721 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt 3781 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc 3841 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata 3901 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac 3961 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg 4021 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt 4081 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt 4141 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc 4201 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga 4261 cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt cgattccacc 4321 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4381 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 4441 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 4501 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 4561 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 4621 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 4681 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 4741 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 4801 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 4861 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 4921 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4981 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 5041 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 5101 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 5161 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 5221 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 5281 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 5341 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 5401 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 5461 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 5521 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 5581 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 5641 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 5701 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 5761 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 5821 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 5881 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 5941 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 6001 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 6061 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 6121 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 6181 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 6241 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 6301 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 6361 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 6421 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 6481 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 6541 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 6601 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 6661 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 6721 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat 6781 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg 6841 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag 6901 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg 6961 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa 7021 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa 7081 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 7141 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 7201 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 7261 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 7321 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 7381 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 7441 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 7501 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 7561 gcagagctct ccctatcagt gatagagatc tccctatcag tgatagagat cgtcgacgag 7621 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata 7681 gaagacaccg ggaccgatcc agcctccgga ctctagcgtt taaactta //