LOCUS V032459 7797 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032459 VERSION V032459 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 927..2534 /label="GBA1(NM_000157)" /note="GBA1(NM_000157)" /gene="GBA1" CDS 2535..2600 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2640..3359 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" polyA_signal 3397..3621 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3667..4095 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4109..4438 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4505..5299 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5473..5594 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5643..5659) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5667..5683 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(5691..5721) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5736..5757 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6045..6630) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6801..7661) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(7662..7766) /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 ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaagcc actgggatcc gccaccatgg agttttcaag tccttccaga gaggaatgtc 961 ccaagccttt gagtagggta agcatcatgg ctggcagcct cacaggattg cttctacttc 1021 aggcagtgtc gtgggcatca ggtgcccgcc cctgcatccc taaaagcttc ggctacagct 1081 cggtggtgtg tgtctgcaat gccacatact gtgactcctt tgaccccccg acctttcctg 1141 cccttggtac cttcagccgc tatgagagta cacgcagtgg gcgacggatg gagctgagta 1201 tggggcccat ccaggctaat cacacgggca caggcctgct actgaccctg cagccagaac 1261 agaagttcca gaaagtgaag ggatttggag gggccatgac agatgctgct gctctcaaca 1321 tccttgccct gtcaccccct gcccaaaatt tgctacttaa atcgtacttc tctgaagaag 1381 gaatcggata taacatcatc cgggtaccca tggccagctg tgacttctcc atccgcacct 1441 acacctatgc agacacccct gatgatttcc agttgcacaa cttcagcctc ccagaggaag 1501 ataccaagct caagataccc ctgattcacc gagccctgca gttggcccag cgtcccgttt 1561 cactccttgc cagcccctgg acatcaccca cttggctcaa gaccaatgga gcggtgaatg 1621 ggaaggggtc actcaaggga cagcccggag acatctacca ccagacctgg gccagatact 1681 ttgtgaagtt cctggatgcc tatgctgagc acaagttaca gttctgggca gtgacagctg 1741 aaaatgagcc ttctgctggg ctgttgagtg gatacccctt ccagtgcctg ggcttcaccc 1801 ctgaacatca gcgagacttc attgcccgtg acctaggtcc taccctcgcc aacagtactc 1861 accacaatgt ccgcctactc atgctggatg accaacgctt gctgctgccc cactgggcaa 1921 aggtggtact gacagaccca gaagcagcta aatatgttca tggcattgct gtacattggt 1981 acctggactt tctggctcca gccaaagcca ccctagggga gacacaccgc ctgttcccca 2041 acaccatgct ctttgcctca gaggcctgtg tgggctccaa gttctgggag cagagtgtgc 2101 ggctaggctc ctgggatcga gggatgcagt acagccacag catcatcacg aacctcctgt 2161 accatgtggt cggctggacc gactggaacc ttgccctgaa ccccgaagga ggacccaatt 2221 gggtgcgtaa ctttgtcgac agtcccatca ttgtagacat caccaaggac acgttttaca 2281 aacagcccat gttctaccac cttggccact tcagcaagtt cattcctgag ggctcccaga 2341 gagtggggct ggttgccagt cagaagaacg acctggacgc agtggcactg atgcatcccg 2401 atggctctgc tgttgtggtc gtgctaaacc gctcctctaa ggatgtgcct cttaccatca 2461 aggatcctgc tgtgggcttc ctggagacaa tctcacctgg ctactccatt cacacctacc 2521 tgtggcgtcg ccaggactac aaagaccatg acggtgatta taaagatcat gacatcgact 2581 acaaggatga cgatgacaag gactcgagtg gcggtggagg aagcggcgga ggtggatcta 2641 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 2701 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 2761 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 2821 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 2881 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 2941 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 3001 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 3061 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 3121 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 3181 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 3241 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 3301 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtagt 3361 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat 3421 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc 3481 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg 3541 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg 3601 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt 3661 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 3721 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 3781 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 3841 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 3901 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 3961 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 4021 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 4081 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 4141 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 4201 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 4261 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 4321 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc 4381 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 4441 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt 4501 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 4561 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 4621 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 4681 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 4741 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 4801 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 4861 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 4921 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 4981 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 5041 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 5101 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 5161 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 5221 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 5281 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc 5341 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 5401 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 5461 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 5521 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 5581 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 5641 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5701 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5761 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5821 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 6001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 6061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 6121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 6241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag 6601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 6661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 6721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 6781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 6841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 6901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 6961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 7021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 7081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 7141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 7201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 7261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 7321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 7381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 7441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 7501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 7561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 7621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 7681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 7741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //