LOCUS V041209 7247 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041209 VERSION V041209 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 923..1639 /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 1676..2737 /label="Aim2(NM_001013779)" /note="Aim2(NM_001013779)" /gene="Aim2" CDS 2744..2809 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2847..3071 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3117..3545 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3559..3888 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3955..4749 /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 4923..5044 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5093..5109) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5117..5133 /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(5141..5171) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5186..5207 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5495..6080) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6251..7111) /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(7112..7216) /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 aagcttgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 1081 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 1321 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 1381 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 1441 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 1621 catggacgag ctgtacaagg gcggtggagg cagtggtgga ggcggtagcg aattcatgga 1681 gagtgagtac cgggaaatgc tgttgttgac cggcctggac cacatcacgg aggaagaact 1741 gaaacggttc aagtactttg ctttgactga gtttcagatt gccaggagca cactcgacgt 1801 ggcagatagg acagagttag ctgaccacct gattcaaagt gcaggtgcgg cgtctgcagt 1861 gaccaaggcc attaatattt tccagaagtt gaattatatg catattgcaa atgctcttga 1921 agagaaaaag aaagaagctg aacgtaaact catgaccaat acaaagaaga gaggaacaca 1981 gaaggtagaa aatagaagtc aagctgaaaa ctgctctgct gcctctgcca cccgcagtga 2041 caatgacttt aaggaacagg ctgctacaga agtctgtcct caagctaagc ctcagaagaa 2101 acagatggtg gcagaacagg aagccatcag agaagattta cagaaagatc cacttgttgt 2161 cacggtgctg aaagctataa atccctttga gtgtgagact caggaaggaa gacaagagat 2221 atttcatgca acagtggcca cggagacaga ttttttcttt gtaaaagttt taaacgcaca 2281 gtttaaagat aaatttatcc caaagaggac aattaaaata tcaaactacc tttggcacag 2341 taacttcatg gaggtcacca gttcctcagt tgtggttgat gttgaatcta accacgaagt 2401 cccaaataac gttgttaaga gagccaggga aactcccagg attagtaaac tgaagattca 2461 gccatgtgga acaattgtga atgggctgtt taaagtccag aagataacag aggaaaaaga 2521 tagagtactg tatggtatac atgataaaac agggacaatg gaggtgttgg tgctgggaaa 2581 cccaagcaaa acaaagtgcg aggaaggaga caagattaga ctcacgttct ttgaggtgtc 2641 aaaaaatgga gtgaaaattc agttgaaatc tggaccttgt agctttttta aggttattaa 2701 ggctgcaaag ccaaaaactg acatgaaaag tgtggagctc gaggactaca aagaccatga 2761 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 2821 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 2881 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2941 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3001 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 3061 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 3121 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 3181 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 3241 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 3301 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 3361 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 3421 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 3481 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 3541 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 3601 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 3661 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 3721 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3781 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 3841 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 3901 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 3961 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 4021 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 4081 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 4141 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 4201 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 4261 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 4321 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 4381 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 4441 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 4501 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 4561 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 4621 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 4681 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4741 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4801 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4861 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4921 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4981 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 5041 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 5101 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 5161 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 5221 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 5281 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 5341 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 5401 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 5461 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 5521 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 5581 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5641 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 5701 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5761 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5821 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5881 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 5941 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 6001 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 6061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 6121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 6181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 6241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 6301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 6361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 6421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 7021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 7081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 7141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 7201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //