LOCUS V037876 6997 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037876 VERSION V037876 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..2487 /label="Hars1(NM_001025414)" /note="Hars1(NM_001025414)" /gene="Hars1" CDS 2494..2559 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2597..2821 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2867..3295 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3309..3638 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3705..4499 /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 4673..4794 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4843..4859) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4867..4883 /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(4891..4921) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4936..4957 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5245..5830) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6001..6861) /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(6862..6966) /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 accatggcag atcgtgctgc gctggaggag ctggtgcgac tccagggagc gcatgtgagg 1021 ggccttaagg agcagaaagc gtccgctgag cagatcgagg aggaggtgac gaaactccta 1081 aaactgaagg cacagctggg ccatgatgaa ggcaaacaga agttcgtcct caaaaccccc 1141 aagggcacga gagactatag tccccggcag atggcagtcc gagaaaaggt gtttgatgtc 1201 atcatccgct gtttcaaacg ccatggagca gaagtgatcg atacacctgt gtttgaacta 1261 aaggagacac taactggaaa atacggagaa gactcaaagc ttatctatga cctgaaggac 1321 cagggtgggg agctgctgtc cctccgttat gacctcactg tcccttttgc tcggtatcta 1381 gcaatgaata aactgaccaa cattaaacgc taccacatag caaaggtgta tcgccgggac 1441 aacccagcta tgacccgagg ccgttaccgt gaattctatc agtgtgattt tgacattgct 1501 ggacagttcg accccatgat tcctgatgca gagtgcctaa agattatgtg tgagatcctg 1561 agttcacttc agattggcaa cttccaggtc aaggtaaatg ataggcgcat cctagacggg 1621 atgtttgctg tctgtggtgt ccctgatagc aagttccgta ccatctgctc ctccgtggac 1681 aagctagaca aggtttcctg ggaggaagtg aagaatgaga tggtgggaga gaagggcctc 1741 gcacctgaag tggctgatcg catcggggac tatgtccagc agcatggtgg ggtgtctctg 1801 gtggaacagt tactccagga tcctaaactc tcccaaaaca aacaggctgt agagggcttg 1861 ggagacctaa agctgctctt tgagtatcta accctatttg gcattgatga caagatctcc 1921 tttgacttga gccttgctcg agggctggac tactacactg gggtcatcta tgaggcagtg 1981 ctgctacaaa tgccaaccca agctggggaa gagcccttgg gtgtgggcag catagctgct 2041 ggagggcgct acgatggact agtaggcatg tttgacccca aagggcgcaa ggtgccatgt 2101 gttgggctca gtattggagt ggaacggatc ttctccattg tagagcaaaa attagaggct 2161 tcagaggaaa aggtgcggac cacagagacg caggtgcttg tggcatctgc acagaagaag 2221 ctgctggagg agagactgaa gcttatctca gagttatggg atgctgggat caaggctgag 2281 ctgctgtaca agaagaaccc aaagttactg aaccagttgc agtactgtga ggaagcaggc 2341 atcccactgg tggccatcat tggtgagcag gagcttaagg atggtgtcat caagctccgt 2401 tcagtgacga gcagagaaga ggtggatgtc cgaagagaag accttgtgga ggagatcaga 2461 agaagaacaa gtcagcccct ctccatgctc gaggactaca aagaccatga cggtgattat 2521 aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc cgtttaaacc 2581 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2641 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2701 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2761 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2821 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2881 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2941 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 3001 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 3061 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3121 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3181 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 3241 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 3301 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3361 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3481 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3541 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3601 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3661 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3721 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3781 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3841 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3901 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3961 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 4021 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4081 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4141 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4201 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4261 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4321 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4381 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4441 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4501 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4561 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4621 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4681 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4741 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4801 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4861 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 4921 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 4981 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 5041 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5101 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5161 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5221 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5281 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5341 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5401 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5461 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5581 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5641 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 5701 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5761 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 6001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6061 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6961 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //