LOCUS V024223 7865 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024223 VERSION V024223 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 935..1651 /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" CDS 1658..3355 /label="Mcoln2(NM_026656)" /note="Mcoln2(NM_026656)" /gene="Mcoln2" CDS 3362..3427 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3465..3689 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3735..4163 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4177..4506 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4573..5367 /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 5541..5662 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5711..5727) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5735..5751 /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(5759..5789) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5804..5825 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6113..6698) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6869..7729) /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(7730..7834) /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 gtttaaactt aagcttggta ccgagctcgg atccatggtg agcaagggcg aggagctgtt 961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 1021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg 1081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt 1141 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 1201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 1261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 1321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 1381 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg 1441 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat 1501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag 1561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 1621 gatcactctc ggcatggacg agctgtacaa ggaattcatg cccggagacg aggagacgct 1681 ggacctgcct gcgtggaacc gcgtcccgga cctgacttgg ggtccccacc acagatctgc 1741 gatggcgtct ctcgattctg aggtgaggga agagtgtctg agggaagacc tgaagttcta 1801 cttcatgagt ccctgtgaga agtaccgagc cagacgtcag atcccgtgga aactgggctt 1861 gcagatattg aagatagtca tggtaaccac acagcttgtt cggtttggct taagtaacca 1921 gttggtggtg gctttcaaag aagataacac tgttgctttt aagcacttgt ttttgaaagg 1981 attttctgga gttgatgaag atgactacag ctgtagcata tacactcagg agaacaccta 2041 tgagagcatc ttttttgcta taaagcagta tcgtcatctg aagaatattt ccctggcaac 2101 ccttggttat ggagaaagtg aagacaatag aactggcttg aaggtctgta agcagcacta 2161 caagacaggg gccatgtttt cttctaacga gaccctgaac attgacagtg acattgagac 2221 agactgcatt cacctggacc ttcaggtgct caccacggaa cccgaggact gggcacagac 2281 atctttcttc aggctggatt tttatcggct cgtgcaagtt gatatctcct ttgccctcaa 2341 aggaattgac ctacaggcag ttcattcccg agagataccg gactgttacc tctttcagaa 2401 tacgattacc ttcgacaaca cagcccacag tgggaaaatc aaaatctatc tcaacagcga 2461 ggccaacatc gaagagtgta aaaacatgaa catatctggg tctactcaga gaagcactca 2521 ctacctcctg gtgtttgatg tgttcgtcat tatgatctgc ctggcatccc ttatcctgtg 2581 cacgagatcc attgttcttg ctctgaggct gaggaagaga tttctaaact tcttcctgga 2641 gaagtacaaa cagcgcgtgt gtggcgctga ccagtgggaa ttcgtcaacg gatggtacgt 2701 cctggtcact atcagcgacc tcatgacaat aattggatcc atattgaaga tggaaatcaa 2761 agccaagaag ctcacaaact atgacgtgtg cagcatcttg ctcgggacgt cgacgctctt 2821 tgtctgggtt ggagtcatca gatatctggg ttactttcag acgtacaacg tgctcatttt 2881 aaccatgcaa gcctcactgc ccaaagttct ccggttctgt gcttgcgctg gcatgatcta 2941 cctggggtac accttctgcg gctggattgt cttaggaccg taccacgaga agtttgaaaa 3001 cctgaacata gtcgctgaat gcctgttttc tctggtcaac ggcgatgaca tgtttgccac 3061 gttcgctcag atccagcaga agagcattct ggtgtggctg ttcagccgcc tctacctgta 3121 ttcctttatc agcctattca tttacatggt cctcagcctc ttcatcgcgc tcatcaccga 3181 ctcttaccac accattaaga agtaccagca gcatgggttt cctgaaacag atctgcagaa 3241 attcctgaag gagagcggaa gcaaagatgg gtaccagaag cagccatcgg ccttgctgtc 3301 ctgcctctgc tgtctgagga ggagaagaag caatgaccac ttgattctca ttgacctcga 3361 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga 3421 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg 3481 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 3541 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 3601 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 3661 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 3721 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 3781 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 3841 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 3901 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 3961 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 4021 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 4081 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 4141 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag 4201 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 4261 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 4321 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 4381 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 4441 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 4501 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga 4561 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 4621 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 4681 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 4741 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 4801 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 4861 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 4921 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 4981 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 5041 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 5101 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 5161 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 5221 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 5281 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 5341 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 5401 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 5461 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 5521 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 5581 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 5641 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 5701 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 5761 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 5821 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 5881 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 5941 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 6001 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 6061 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 6121 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 6181 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 6241 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 6301 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 6361 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 6421 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 6481 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 6541 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 6601 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt 6661 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 6721 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 6781 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 6841 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 6901 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 6961 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 7021 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 7081 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 7141 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 7201 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 7261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 7321 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 7381 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 7441 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 7501 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 7561 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 7621 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 7681 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 7741 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7801 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7861 acgtc //