LOCUS V043696 6784 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043696 VERSION V043696 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..2274 /label="Plin3(NM_025836)" /note="Plin3(NM_025836)" /gene="Plin3" CDS 2281..2346 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2384..2608 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2654..3082 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3096..3425 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3492..4286 /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 4460..4581 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4630..4646) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4654..4670 /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(4678..4708) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4723..4744 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5032..5617) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5788..6648) /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(6649..6753) /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 accatgtcta gcaatggtac agatgcgcct gccgaagctc aagctgctat ggaggaacct 1021 gttgtgcagc ccagcgtggt ggatcgtgtt gccggcctgc ctcttatcag ttccacgtat 1081 gggatggtga gcgccgccta cacctccacc aaggagaact acccccacgt caggactgtg 1141 tgcgatgtgg ctgagaaagg cgtcaagacc ctcaccacgg ccgctgtcag cacagcacag 1201 cccatcctgt ccaagctgga gccccagatt gcgacggcca gtgagtatgc acaccgaggg 1261 ctggacagac tgcaggaaag cctgcccatc ctccagcagc cgactgagaa ggttttggcg 1321 gatactaagg aactggtgtc atctacagtg tctggggctc aagaaatggt atctagctca 1381 gtgtctagtg cgaaggaaac agtggccacc cgggtcacag gagcggtgga tgtgaccctc 1441 ggggctgtgc agaacagcgt ggacaagacc aagtcagcaa tgaccagcgg tgtccaatca 1501 gtcatgggct cacgtgtggg acagatggtg attagcggag tggacagagt gcttgtgaaa 1561 tcagaggcct gggcagacaa tcgcctgccc ctgacagagg cggagctagc tctgattgcc 1621 acacccccgg aggactcgga catggcctca ctgcagcagc agcgacagga gcagaactac 1681 tttgtccggt tgggctctct ctctgagagg ctgcgcaatc acgcctatga acactccctc 1741 ggcaagctgc agaatgccag gcagaaggca caggagacgc tgcagcagct aacaagtgtg 1801 ctcggcctga tggaatccgt gaaacagggt gtggaccaga ggctggggga ggggcaggag 1861 aagctgcacc agatgtggct cagctggaac cagaagaccc cacaggatgc tgaaaaggac 1921 ccagctaagc cagagcaggt ggaggcccgg gcactcagca tgttcaggga catcacccag 1981 cagctgcaga gcatgtgtgt ggccctgggg gccagcatcc agggcttgcc cagccacgtc 2041 agggagcagg cacagcaggc ccgcagccag gtgaacgacc ttcaggccac cttctccggc 2101 attcactcct tccaggacct ctcagccggc gttcttgccc aaactcggga gcgcatagcc 2161 agagcccggg aggccctaga caacactgtg gagtacgtgg cccagaacac tccagccatg 2221 tggctggtgg gccccttcgc tcctggaatc acagagaaaa cccctgaagg gaagctcgag 2281 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat 2341 gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 2401 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 2461 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 2521 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 2581 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 2641 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2701 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2761 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2821 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 2881 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 2941 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3001 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 3061 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 3121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3361 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3421 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3481 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3541 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3601 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3661 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3721 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 3781 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 3841 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 3901 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 3961 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4021 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4081 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4141 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4201 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4261 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 4321 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 4381 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 4441 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 4501 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 4561 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 4621 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 4681 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 4741 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 4801 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 4861 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4921 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4981 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5041 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5101 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 5161 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 5221 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 5281 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 5341 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 5401 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5461 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5521 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 5581 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 5641 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 5701 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 5761 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 5821 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 5881 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 5941 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 6001 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 6061 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 6121 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 6181 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 6241 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 6301 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 6361 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 6421 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 6481 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 6541 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 6601 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 6661 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 6721 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 6781 cgtc //