LOCUS V039465 7765 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039465 VERSION V039465 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..3255 /label="Sarm1(NM_001168521)" /note="Sarm1(NM_001168521)" /gene="Sarm1" CDS 3262..3327 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3365..3589 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3635..4063 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4077..4406 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4473..5267 /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 5441..5562 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5611..5627) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5635..5651 /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(5659..5689) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5704..5725 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6013..6598) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6769..7629) /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(7630..7734) /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 accatggtcc tgacgctgct cttctccgcc tacaaactgt gccgcttctt caccatgtca 1021 ggcccacggc cgggcgccga tcggctgaca gtgcccggac cggatcggag tggtggcgcc 1081 agcccatggt gggctgcggg cggtcgcggg tctcgcgaag tgtcgcccgg agtgggcact 1141 gaggtgcaag gcgccctgga gcgttcgctg cctgagctgc agcaggcgct gtccgagctg 1201 aaacaggcaa gcgcggcgcg ggctgtgggc gcgggtctcg ccgaggtctt ccagctggta 1261 gaggaagcct ggctgctgcc ggccgtgggc cgcgaggtgg cccaaggtct atgcgatgct 1321 atacgtctgg acggtggcct cgacttgctg ttgcggctgc ttcaggcacc ggagctagag 1381 acccgtgtgc aggccgcgcg cttgctggag cagatcctgg tggctgagaa ccgggaccgc 1441 gtggcgcgca tcggtctagg cgtgatcttg aacctggcga aggagcgcga gcctgtggaa 1501 ctggcacgaa gcgtggcggg catcttggag cacatgttca agcactcgga ggagacgtgc 1561 cagcggctgg tggcggccgg aggcctcgac gcggtgctgt actggtgccg ccgcacagac 1621 ccggcgctgc tgcgccactg cgctcttgcg ctggcgaact gcgcgctgca cgggggccag 1681 acggtgcagc ggtgcatggt ggagaagcgc gccgccgagt ggctcttccc gctcgctttc 1741 tccaaggagg acgagctgct gcggctgcac gcctgcctgg cggtggcggt gttggctacc 1801 aacaaggagg tggaacgcga ggtcgagcat tctggcacat tggcgcttgt cgagccgctc 1861 gtggcatcgc tggaccccgg ccgcttcgcc cgctgcctgg tggatgccag tgacacaagc 1921 cagggtcgtg gaccagacga cctgcagagc ctggtgctgt tgctcgattc gtcgcgtttg 1981 gaggctcagt gcataggagc attctacctg tgcgcagagg ctgccatcaa gagcctacag 2041 ggaaagacca aggtgttcag cgacatcggc gctatccaga gcctgaaacg cctggtttct 2101 tactctacga atggcaccac gtcggcgctg gccaagcgcg cgctgcgcct attgggcgag 2161 gaggtgccaa ggcgcatcct gccctgcgtg gccagctgga aggaagctga ggtccagacc 2221 tggctacagc agatcggctt ctcccagtac tgcgagaact ttcgggagca gcaggtagat 2281 ggtgacctgc ttctaagact cacagatgaa gaactccaga cagacctagg catgaaatca 2341 agcatcaccc gcaagaggtt ctttagggag ctcacagagc tcaagacctt cgccagctac 2401 gctacttgcg accgcagcaa cctagcggac tggctgggca gcctggatcc tcgcttccgc 2461 cagtacacct atggcctggt cagctgcggt ctggaccgct ccctgctgca ccgcgtgtca 2521 gagcagcagc tcctggagga ctgtggcatc cgcctgggag tgcaccgcac gcgcatcctc 2581 tctgcagcca gaggtcactt tgcccagact ggcctgagaa gcttgaggag accaagtctc 2641 cacgatgatg gaccccgtga taagcagtgg ggaagagcca ccctcacctc catgtctctt 2701 tccttggctc cagaaatgct acattccccg ctgccctgta ctggaggcaa gctcagtggg 2761 gacaccccag atgtctttat cagttaccgg aggaactcag ggtcccagct ggccagcctc 2821 ctgaaggtgc acctgcagct tcacggcttc agcgtcttca tcgacgtgga gaagctggaa 2881 gccggcaaat tcgaggacaa gcttatccaa agcgtcatag cggctcgcaa ttttgtcctg 2941 gtgctgtctg ctggggcgct ggataagtgc atgcaggacc atgactgcaa ggactgggtg 3001 cacaaggaga ttgtgactgc tttaagctgt ggcaagaaca ttgtgcccat cattgatggc 3061 tttgagtggc ctgagcctca ggcgctgcct gaggatatgc aggctgtact caccttcaac 3121 ggcatcaaat ggtcccatga gtaccaggag gccaccatcg agaagatcat ccgcttccta 3181 cagggccgcc cctctcagga ctcctctgcc ggatcggata ccagtttgga gggagctacg 3241 ccaatgggtc tgcctctcga ggactacaaa gaccatgacg gtgattataa agatcatgac 3301 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc 3361 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 3421 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 3481 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 3541 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 3601 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 3661 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3721 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3781 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3841 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3901 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3961 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 4021 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 4081 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 4141 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 4201 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 4261 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 4321 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 4381 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 4441 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 4501 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 4561 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 4621 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 4681 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 4741 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 4801 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 4861 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 4921 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 4981 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 5041 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 5101 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 5161 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 5221 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 5281 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 5341 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 5401 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 5461 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 5521 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 5581 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 5641 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 5701 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5761 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 5821 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 5881 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 5941 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6001 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6061 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6181 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 6421 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 6481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 6541 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 6601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 6721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6901 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 7621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 7681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 7741 tccccgaaaa gtgccacctg acgtc //