LOCUS V027039 8024 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027039 VERSION V027039 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..3514 /label="EEF2(NM_001961)" /note="EEF2(NM_001961)" /gene="EEF2" CDS 3521..3586 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3624..3848 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3894..4322 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4336..4665 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4732..5526 /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 5700..5821 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5870..5886) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5894..5910 /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(5918..5948) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5963..5984 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6272..6857) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7028..7888) /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(7889..7993) /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 atggtgaact tcacggtaga 961 ccagatccgc gccatcatgg acaagaaggc caacatccgc aacatgtctg tcatcgccca 1021 cgtggaccat ggcaagtcca cgctgacaga ctccctggtg tgcaaggcgg gcatcatcgc 1081 ctcggcccgg gccggggaga cacgcttcac tgatacccgg aaggacgagc aggagcgttg 1141 catcaccatc aagtcaactg ccatctccct cttctacgag ctctcggaga atgacttgaa 1201 cttcatcaag cagagcaagg acggtgccgg cttcctcatc aacctcattg actcccccgg 1261 gcatgtcgac ttctcctcgg aggtgactgc tgccctccga gtcaccgatg gcgcattggt 1321 ggtggtggac tgcgtgtcag gcgtgtgcgt gcagacggag acagtgctgc ggcaggccat 1381 tgccgagcgc atcaagcctg tgctgatgat gaacaagatg gaccgcgccc tgctggagct 1441 gcagctggag cccgaggagc tctaccagac tttccagcgc atcgtggaga acgtgaacgt 1501 catcatctcc acctacggcg agggcgagag cggccccatg ggcaacatca tgatcgatcc 1561 tgtcctcggt accgtgggct ttgggtctgg cctccacggg tgggccttca ccctgaagca 1621 gtttgccgag atgtatgtgg ccaagttcgc cgccaagggg gagggccagt tggggcctgc 1681 cgagcgggcc aagaaagtag aggacatgat gaagaagctg tggggtgaca ggtactttga 1741 cccagccaac ggcaagttca gcaagtcagc caccagcccc gaagggaaga agctgccacg 1801 caccttctgc cagctgatcc tggaccccat cttcaaggtg tttgatgcga tcatgaattt 1861 caagaaagag gagacagcaa aactgataga gaaactggac atcaaactgg acagcgagga 1921 caaggacaaa gaaggcaaac ccctgctgaa ggctgtgatg cgccgctggc tgcctgccgg 1981 agacgccttg ttgcagatga tcaccatcca cctgccctcc cctgtgacgg cccagaagta 2041 ccgctgcgag ctcctgtacg aggggccccc ggacgacgag gctgccatgg gcattaaaag 2101 ctgtgacccc aaaggccctc ttatgatgta tatttccaaa atggtgccaa cctccgacaa 2161 aggtcggttc tacgcctttg gacgagtctt ctcggggctg gtctccactg gcctgaaggt 2221 caggatcatg gggcccaact atacccctgg gaagaaggag gacctctacc tgaagccaat 2281 ccagagaaca atcttgatga tgggccgcta cgtggagccc atcgaggatg tgccttgtgg 2341 gaacattgtg ggcctcgtgg gcgtggacca gttcctggtg aagacgggca ccatcaccac 2401 cttcgagcac gcgcacaaca tgcgggtgat gaagttcagc gtcagccctg ttgtcagagt 2461 ggccgtggag gccaagaacc cggctgacct gcccaagctg gtggaggggc tgaagcggct 2521 ggccaagtcc gaccccatgg tgcagtgcat catcgaggag tcgggagagc atatcatcgc 2581 gggcgccggc gagctgcacc tggagatctg cctgaaggac ctggaggagg accacgcctg 2641 catccccatc aagaaatctg acccggtcgt ctcgtaccgc gagacggtca gtgaagagtc 2701 gaacgtgctc tgcctctcca agtcccccaa caagcacaac cggctgtaca tgaaggcgcg 2761 gcccttcccc gacggcctgg ccgaggacat cgataaaggc gaggtgtccg cccgtcagga 2821 gctcaagcag cgggcgcgct acctggccga gaagtacgag tgggacgtgg ctgaggcccg 2881 caagatctgg tgctttgggc ccgacggcac cggccccaac atcctcaccg acatcaccaa 2941 gggtgtgcag tacctcaacg agatcaagga cagtgtggtg gccggcttcc agtgggccac 3001 caaggagggc gcactgtgtg aggagaacat gcggggtgtg cgcttcgacg tccacgacgt 3061 caccctgcac gccgacgcca tccaccgcgg agggggccag atcatcccca cagcacggcg 3121 ctgcctctat gccagtgtgc tgaccgccca gccacgcctc atggagccca tctaccttgt 3181 ggagatccag tgtccagagc aggtggtcgg tggcatctac ggggttttga acaggaagcg 3241 gggccacgtg ttcgaggagt cccaggtggc cggcaccccc atgtttgtgg tcaaggccta 3301 tctgcccgtc aacgagtcct ttggcttcac cgctgacctg aggtccaaca cgggcggcca 3361 ggcgttcccc cagtgtgtgt ttgaccactg gcagatcctg cccggagacc ccttcgacaa 3421 cagcagccgc cccagccagg tggtggcgga gacccgcaag cgcaagggcc tgaaagaagg 3481 catccctgcc ctggacaact tcctggacaa attgctcgag gactacaaag accatgacgg 3541 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 3601 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 3661 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 3721 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 3781 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 3841 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 3901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 3961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 4021 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 4081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 4141 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 4201 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 4261 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 4321 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 4381 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 4441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 4501 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 4561 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 4621 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 4681 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 4741 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 4801 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 4861 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 4921 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 4981 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 5041 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 5101 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 5161 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 5221 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 5281 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 5341 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 5401 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 5461 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 5521 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 5581 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 5641 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 5701 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 5761 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 5821 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 5881 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 5941 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 6001 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 6061 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 6121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 6181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 6241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 6301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 6361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 6421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 6481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 6541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 6601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 6661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 6721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 6781 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 6841 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 6901 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 6961 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 7021 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 7081 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 7141 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 7201 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 7261 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 7321 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 7381 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 7441 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 7501 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 7561 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 7621 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 7681 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 7741 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 7801 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 7861 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 7921 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 7981 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //