LOCUS V019467 7094 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019467 VERSION V019467 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 941..2626 /label="Gmeb1(NM_020273)" /note="Gmeb1(NM_020273)" /gene="Gmeb1" CDS 2633..2656 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" polyA_signal 2694..2918 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2964..3392 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3406..3735 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3802..4596 /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 4770..4891 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4940..4956) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4964..4980 /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(4988..5018) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5033..5054 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5342..5927) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6098..6958) /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(6959..7063) /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 atccgccacc atggccaatg cagaagtgag 961 cgtcccagtg ggagatgtgg ttgtggtacc cactgaagga aatgaagggg agaacccgga 1021 agacactaaa actcaagtga tcttgcagtt acagcctgtg caacaaggga tttatgaagc 1081 tgggtcggag aatagcgcag cagttgtagc cgtagagacc cactcaatac acaaaatcga 1141 ggaaggaatt gatgcaagca gtatagaagg gaacgaggac atggagatcg cttaccctat 1201 aacctgtggg gagagtaaag ctgtcctcct ctggaagaag ttcgtgtgtc caggaataaa 1261 cgtgaagtgt gtcaagttta atgaccagct gatcagtccc aagcacttcg ttcacctggc 1321 tggcaagtcc accctgaagg actggaagag agccatccgt ctaggcggca tcatgctcag 1381 gaagatgatg gactctggac agatcgattt ttaccagcat gacaaagttt gctccaatac 1441 ctgcaggagc accaaatttg acctcctgat cagcagtgcg agggctccag tgccaggaca 1501 gcagacaagt gtggtgcaga cccccacctc ggccgatggg aacatcacac agatcgccat 1561 ctcagaggag agcatggaag aggcagggct ggagtggaac tcagctctca ccgccgctgt 1621 caccatggcc acagaggagg gtataaagaa agagtctgaa gagatttcag aggacacttt 1681 gatgttctgg aaggggatag ctgatgtagg gttgatggag gaggtcgtct gcaacatcca 1741 gaaggagatg gaagagctgc tcaggggcgt tcagcagcgg ctcattcagg cccctttcca 1801 ggtcacagat gctgctgttc taaacaatgt ggcgaacaca tttggcctaa tggacgcggt 1861 caagagagtt ttggacaaca gacggaagca agtggagcag ggcgaggagc agtttctcta 1921 caccctggca gacttggaac ggcagttgga ggagcaaaaa aagcaagccc aagatcccag 1981 actgaaatcc cagacagttc aaaatgtggt actgatgccc gtgagcaccc caaagcctcc 2041 aaaaagaccc cggctccagc ggccagcctc caccaccgtc ctgagccctt ctcccgtcca 2101 gcagcctcag tttactgtca tctctcccat caccattact cctgtgggcc agtccttctc 2161 catgggcaac atcccagtgg ctaccctcag ccagggctcc agtcctgtga ctgtccacac 2221 actgccttct ggccctcagc tcttccgcta tgccacggtg gtttcctctg ccaagagcaa 2281 ctcaccagac acagtgacca tccacccttc ctcaagtttg gcactgctaa gctccacctc 2341 tatgcaggat gggagttccc tgggcaacat ggccaccatg gtgagcccca tggagctggt 2401 ggccatggag tccggcctga cgtcagcaat ccaggctgtt gaaagcacct ctgaggatgg 2461 gcagaccatc attgaaatcg accctgcccc agactccgag gctgacgaca ctgagggcaa 2521 agcagtcatc ttggagacag ggctgaggac tgaggagaaa gtggtagccg aaatggaaga 2581 acaccagcat caggtccaca atgtggagat tgtggtcttg gaggacctcg aggattacaa 2641 ggatgacgac gataagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 2701 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 2761 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 2821 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 2881 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 2941 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 3001 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 3061 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 3121 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 3181 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 3241 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 3301 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 3361 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 3421 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3481 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3541 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 3601 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 3661 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 3721 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 3781 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 3841 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 3901 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 3961 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 4021 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 4081 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 4141 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 4201 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 4261 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 4321 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 4381 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 4441 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 4501 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 4561 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 4621 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 4681 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 4741 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 4801 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 4861 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 4921 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 4981 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 5041 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 5101 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 5161 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 5221 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 5281 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 5341 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 5401 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5461 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5521 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5581 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 5641 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5701 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5761 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 5821 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt 5881 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 5941 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 6001 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 6061 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 6121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 6181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 6241 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 6301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 6361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 6421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 6481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 6541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 6601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 6661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 6721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 6781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 6841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 6901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 6961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 7021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 7081 tgccacctga cgtc //