LOCUS V023094 5867 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023094 VERSION V023094 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..1357 /label="MSMP(NM_001044264)" /note="MSMP(NM_001044264)" /gene="MSMP" CDS 1364..1429 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1467..1691 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1737..2165 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2179..2508 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2575..3369 /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 3543..3664 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3713..3729) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3737..3753 /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(3761..3791) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3806..3827 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4115..4700) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4871..5731) /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(5732..5836) /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 atggccctaa ggatgctctg 961 ggctggacag gccaagggga tcctaggagg ctgggggatc atctgcttgg tgatgtctct 1021 actcctccag cacccaggag tctacagcaa gtgctacttc caagctcaag ccccctgtca 1081 ctatgagggg aaatatttta ccctgggtga gtcttggctc cgcaaggact gtttccattg 1141 cacctgtctg catcctgttg gcgtgggctg ctgtgacacg tcccagcatc ccatcgactt 1201 cccggctggg tgtgaggtac gtcaggaggc aggaacctgc cagttctcct tggtgcaaaa 1261 atctgaccct cggctgccct gcaaaggggg agggcctgac ccagaatggg gctcagccaa 1321 cacccctgtt cctggggctc ctgctcccca ctccagcctc gaggactaca aagaccatga 1381 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 1441 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 1501 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 1561 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 1621 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 1681 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 1741 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 1801 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 1861 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 1921 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 1981 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 2041 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 2101 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 2161 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 2221 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 2281 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 2341 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 2401 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 2461 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 2521 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 2581 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 2641 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 2701 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 2761 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 2821 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 2881 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 2941 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3001 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3061 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 3121 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3181 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3241 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 3301 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 3361 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 3421 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 3481 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 3541 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 3601 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 3661 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 3721 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 3781 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 3841 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 3901 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 3961 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4021 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4081 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4141 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4201 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4261 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4321 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4381 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 4441 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 4501 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 4561 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 4621 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 4681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 4741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 4801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 4861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 4921 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 4981 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 5041 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 5101 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 5161 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 5221 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 5281 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 5341 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 5401 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 5461 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 5521 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 5581 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 5641 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 5701 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 5761 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 5821 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //