LOCUS V040075 7097 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040075 VERSION V040075 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..2587 /label="H1f8(NM_138311)" /note="H1f8(NM_138311)" /gene="H1f8" CDS 2594..2659 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2697..2921 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2967..3395 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3409..3738 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3805..4599 /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 4773..4894 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4943..4959) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4967..4983 /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(4991..5021) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5036..5057 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5345..5930) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6101..6961) /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(6962..7066) /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 aattcatggc 1681 tcctgggagt gtctccagtg tttcttcctc ctcttttccc tccagggaca catccccttc 1741 tggatcatgt gggctccctg gagctgacaa gccaggtcca agttgccgca gaatccaagc 1801 aggccaaagg aacccaacaa tgctgcacat ggtgctagag gctttgaagg cccgggaggc 1861 acgccagggc acatcagttg tagccatcaa ggtctacatc caacacaagt acccgacagt 1921 ggacaccacc cgtttcaagt acctgttgaa gcaagctctg gaaactggcg ttcgtcgagg 1981 cctcctcacc aggcctgctc actccaaggc caagggtgcc actggcagct tcaaactagt 2041 tccaaagccc aagacaaaga aagcctgtgc ccccaaagcc ggcaggggag ctgcaggtgc 2101 caaggagaca ggctccaaga aatctggatt gctgaagaaa gaccaagttg gcaaggccac 2161 gatggagaaa gggcagaaga ggagggctta cccttgcaag gcagccacac tggagatggc 2221 acctaagaaa gccaaggcga aaccgaaaga ggtcagaaag gctcccctaa aacaagacaa 2281 agcagcaggg gcccctctga ctgccaatgg aggccagaag gtcaaacgca gtgggagcag 2341 gcaagaagca aatgcccatg ggaaaaccaa aggtgagaaa tcgaagccct tggccagcaa 2401 ggtccagaat agcgttgctt ccctcgccaa aaggaagatg gcagacatgg cccacactgt 2461 gacagttgtt cagggggctg agacagtaca ggagaccaaa gtgcccactc cttcccagga 2521 cataggacac aaagtacaac ccatacctag ggtcaggaag gcaaagaccc ctgagaacac 2581 tcaggccctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta 2641 caaggatgac gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt 2701 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 2761 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 2821 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 2881 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 2941 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 3001 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3061 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3121 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3181 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 3241 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 3301 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 3361 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 3421 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3481 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3541 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3601 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3661 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3721 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 3781 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3841 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 3901 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 3961 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 4021 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4081 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4141 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4201 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4261 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4321 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4381 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4441 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4501 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4561 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4621 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4681 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4741 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 4801 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4861 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 4921 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 4981 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 5041 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5101 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5761 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 5881 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 5941 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 6001 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 6061 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 6121 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6181 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6241 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6301 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6361 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6421 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6481 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6541 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6601 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6661 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6721 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6781 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6841 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 6901 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 6961 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 7021 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 7081 aagtgccacc tgacgtc //