LOCUS V045076 8287 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045076 VERSION V045076 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 964..3777 /label="Cblb(NM_001033238)" /note="Cblb(NM_001033238)" /gene="Cblb" CDS 3784..3849 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3887..4111 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4157..4585 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4599..4928 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4995..5789 /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 5963..6084 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6133..6149) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6157..6173 /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(6181..6211) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6226..6247 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6535..7120) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7291..8151) /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(8152..8256) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatggcaa attctatgaa tggcagaaat cctggtggtc gaggaggaaa cccccgcaaa 1021 ggtcgcattt tggggattat tgatgctatt caggatgcag tcggaccccc aaagcaagct 1081 gcagctgacc gcaggactgt ggagaagact tggaaactca tggacaaagt ggtaagactg 1141 tgccaaaatc ccaagcttca gttgaaaaac agcccaccgt atatacttga tattttacct 1201 gatacgtatc agcacttgag acttatattg agtaaatatg atgacaacca gaagctggct 1261 caactgagcg agaatgagta ctttaaaatc tacatcgata gtctcatgaa gaagtcgaag 1321 cgagcgatcc ggctctttaa agaaggcaag gaaaggatgt acgaagagca gtcgcaggac 1381 agacggaatc tcacaaagct gtcccttatc ttcagtcaca tgctggcaga aatcaaggcg 1441 atctttccca atggccagtt ccagggagat aacttccgga tcaccaaagc agatgctgct 1501 gagttctgga ggaagttttt tggagacaaa actattgtac catggaaagt cttcagacag 1561 tgcctgcatg aggtccatca gatcagctct ggcctggaag caatggctct gaagtcaacc 1621 attgatttaa cttgcaatga ttacatctca gtgtttgaat ttgatatttt taccaggcta 1681 tttcagccct ggggctctat tttacggaat tggaacttct tggctgtgac tcacccggga 1741 tacatggcat ttctcacgta tgatgaagtt aaagctcggc tacagaaata cagcaccaag 1801 cctggaagtt acattttccg gttaagctgc actcggctgg gacaatgggc cattggctat 1861 gtgactgggg acggcaatat cctacagacc atacctcata acaagcccct gttccaagcc 1921 ctgattgatg gtagcaggga aggcttttat ctttatcctg atgggcggag ctataaccct 1981 gatttaaccg gattatgtga acctacacct cacgatcata taaaagttac acaggagcag 2041 tatgaactgt attgtgaaat gggctccact tttcagctct gcaagatctg tgcagagaat 2101 gacaaagatg tcaagattga gccttgcggg catctgatgt gcacttcgtg ccttaccgcg 2161 tggcaggagt ctgatggcca aggctgcccc ttctgtcgct gtgagataaa aggaacggag 2221 cccatcattg tggacccctt cgaccccaga gatgaaggct ccaggtgctg cagcatcatt 2281 gaccctttca gcatccccat gcttgacttg gacgatgacg atgatcgaga ggagtctttg 2341 atgatgaaca ggctggcgag tgttcgaaag tgcacggaca ggcagaactc accagtcaca 2401 tcgccaggct cctcacccct tgcccagaga agaaagcctc agccagaccc tctccagatc 2461 ccccacctca gcctgccacc agtgcctccc cgcctagatc tcattcagaa aggcatcgtg 2521 cgctctccgt gtggcagccc cacaggctcg ccaaagtctt ctccatgcat ggttagaaaa 2581 caagacaaac cactcccggc accccctcct cccttgagag atccacctcc tccaccagag 2641 aggcctcccc ccatcccacc tgacaataga ctgagcagac acttccacca tggagagagt 2701 gtgccttccc gggaccagcc catgcccctc gaagcctggt gccctcggga tgccttcggg 2761 actaaccagg tgatgggatg tcgcatcctc ggggatggct ctccaaagcc tggcgtcacc 2821 gcaaactcca gcttaaatgg gaggcacagt agaatgggct ccgagcaggt tcttatgagg 2881 aaacacagac gccatgattt gccttcagaa ggagccaagg tcttttccaa tggccacctt 2941 gccactgaag aatacgatgt ccctcctcgg ctttcccctc cgcctccagt caccaccctt 3001 ctcccaagca taaagtgtac tggtccatta gcaaattgtc tctcagagaa aactagagac 3061 acggtagaag atgatgacga tgaatacaag attccttcat cccatcctgt gtccctgaat 3121 tcacaaccat ctcattgcca taatgtcaaa gctcctgttc ggtcttgtga taatggtcat 3181 tgtatattga atggaactca tggtgcgcct tcagagatga agaaatcgaa catcccagat 3241 ttaggcatct atttgaaggg tgaggacgcc tttgatgctc tccctccatc cctccctcct 3301 cccccacctc ctgcaagaca cagtctcatc gaacattcaa aacctccagg ctccagtagc 3361 cggccttcct cagggcagga ccttttcctt cttccttcag atcccttttt tgacccaaca 3421 agtggccaag ttccattgcc tcctgccagg agagcagcag gagacagcgg caaagccaac 3481 agagcctcgc aggactatga ccagctccct tcatcttccg atggttcgca agcaccagct 3541 agacccccca aaccacgacc ccgaaggact gcaccagaaa ttcatcacag aaagccccat 3601 gggcccgaag cggcgttgga aaatgtcgat gcaaaaattg caaaactcat gggagagggg 3661 tatgcctttg aagaggtgaa gagagcctta gagattgccc agaataacgt ggaagtggcc 3721 aggagcatac ttcgagaatt tgccttcccc cctcctgtct ccccacgtct gaatctactc 3781 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3841 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt 3901 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 3961 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 4021 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 4081 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 4141 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 4201 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 4261 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 4321 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 4381 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 4441 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 4501 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 4561 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 4621 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 4681 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 4741 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 4801 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4861 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 4921 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 4981 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 5041 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 5101 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 5161 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 5221 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 5281 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 5341 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 5401 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 5461 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 5521 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 5581 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 5641 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 5701 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 5761 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 5821 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 5881 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 5941 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 6001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 6061 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 6121 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt 7081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 7141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 7201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 7261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 7321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 7381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 7441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 7501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 7561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 7621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 7681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 7741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 8041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 8101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 8161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 8221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 8281 tgacgtc //