LOCUS V038047 6187 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038047 VERSION V038047 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..1677 /label="Dram1(NM_027878)" /note="Dram1(NM_027878)" /gene="Dram1" CDS 1684..1749 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1787..2011 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2057..2485 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2499..2828 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2895..3689 /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 3863..3984 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4033..4049) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4057..4073 /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(4081..4111) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4126..4147 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4435..5020) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5191..6051) /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(6052..6156) /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 accatgctgt gcttcctcag gggaatggct ttcgtcccct tcctcctggt gacttggtcg 1021 tccgcagcct tcatcatctc ctacgtggtc gcggtgctct ctgggcacgt caaccccttt 1081 ctcccctata tcagtgacac aggaacaact cctccagaga gtggtatttt tggattcatg 1141 ataaacttct ctgcatttct tggcgcagct acgatgtaca caagatacaa gatagtggag 1201 aagcagaatg agacctgcta cttcagcact cccgttttta acttggtgtc cttggcgctt 1261 ggattggtgg gatgcatcgg aatgggcatc gtagccaact tccaggagtt agccgtgcct 1321 gtggtccatg atggcggtgc gcttctggct ttcgtctgcg gggtggtgta cacgctcctg 1381 caatcgatca tctcctacaa atcctgtccc cagtggaaca gtctcaccac gtgccatgtc 1441 aggatggcca tctccgctgt ttcgtgcgca gctgtcgtcc ccatgattgc ctgtgcttca 1501 ctcatttcta taaccaagct ggaatggaat ccaaaagaaa aggattatat atatcacgtg 1561 gtgagcgcca tctgtgagtg gaccgtggct tttggtttta ttttctattt cctaacattc 1621 atccaagatt tccagagtgt cactctaagg atatccacag aaatcaatga cgactttctc 1681 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 1741 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt 1801 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 1861 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 1921 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 1981 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 2041 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 2101 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 2161 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 2221 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 2281 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 2341 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 2401 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 2461 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 2521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 2581 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 2641 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 2701 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 2761 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 2821 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 2881 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 2941 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3001 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3061 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 3121 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 3181 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 3241 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 3301 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 3361 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 3421 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 3481 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 3541 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 3601 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 3661 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 3721 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 3781 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 3841 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 3901 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 3961 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 4021 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 4081 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 4141 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 4201 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 4261 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 4321 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 4381 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 4441 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 4501 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 4561 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 4621 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 4681 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 4741 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 4801 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 4861 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 4921 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt 4981 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5041 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 5101 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 5161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 5221 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 5281 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 5341 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 5401 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 5461 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 5521 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 5581 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 5641 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 5701 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 5761 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 5821 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 5881 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 5941 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 6001 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 6061 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 6121 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 6181 tgacgtc //