LOCUS V038073 7069 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038073 VERSION V038073 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 931..2505 /label="LAG3(NM_002286)" /note="LAG3(NM_002286)" /gene="LAG3" CDS 2512..2601 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" polyA_signal 2636..2860 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2906..3334 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3348..3677 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3744..4538 /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 4712..4833 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4882..4898) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4906..4922 /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(4930..4960) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4975..4996 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5284..5869) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6040..6900) /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(6901..7005) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cactagtcca gtgtggtgga attcgccacc atgtgggagg ctcagttcct gggcttgctg 961 tttctgcagc cgctttgggt ggctccagtg aagcctctcc agccaggggc tgaggtcccg 1021 gtggtgtggg cccaggaggg ggctcctgcc cagctcccct gcagccccac aatccccctc 1081 caggatctca gccttctgcg aagagcaggg gtcacttggc agcatcagcc agacagtggc 1141 ccgcccgctg ccgcccccgg ccatcccctg gcccccggcc ctcacccggc ggcgccctcc 1201 tcctgggggc ccaggccccg ccgctacacg gtgctgagcg tgggtcccgg aggcctgcgc 1261 agcgggaggc tgcccctgca gccccgcgtc cagctggatg agcgcggccg gcagcgcggg 1321 gacttctcgc tatggctgcg cccagcccgg cgcgcggacg ccggcgagta ccgcgccgcg 1381 gtgcacctca gggaccgcgc cctctcctgc cgcctccgtc tgcgcctggg ccaggcctcg 1441 atgactgcca gccccccagg atctctcaga gcctccgact gggtcatttt gaactgctcc 1501 ttcagccgcc ctgaccgccc agcctctgtg cattggttcc ggaaccgggg ccagggccga 1561 gtccctgtcc gggagtcccc ccatcaccac ttagcggaaa gcttcctctt cctgccccaa 1621 gtcagcccca tggactctgg gccctggggc tgcatcctca cctacagaga tggcttcaac 1681 gtctccatca tgtataacct cactgttctg ggtctggagc ccccaactcc cttgacagtg 1741 tacgctggag caggttccag ggtggggctg ccctgccgcc tgcctgctgg tgtggggacc 1801 cggtctttcc tcactgccaa gtggactcct cctgggggag gccctgacct cctggtgact 1861 ggagacaatg gcgactttac ccttcgacta gaggatgtga gccaggccca ggctgggacc 1921 tacacctgcc atatccatct gcaggaacag cagctcaatg ccactgtcac attggcaatc 1981 atcacagtga ctcccaaatc ctttgggtca cctggatccc tggggaagct gctttgtgag 2041 gtgactccag tatctggaca agaacgcttt gtgtggagct ctctggacac cccatcccag 2101 aggagtttct caggaccttg gctggaggca caggaggccc agctcctttc ccagccttgg 2161 caatgccagc tgtaccaggg ggagaggctt cttggagcag cagtgtactt cacagagctg 2221 tctagcccag gtgcccaacg ctctgggaga gccccaggtg ccctcccagc aggccacctc 2281 ctgctgtttc tcatccttgg tgtcctttct ctgctccttt tggtgactgg agcctttggc 2341 tttcaccttt ggagaagaca gtggcgacca agacgatttt ctgccttaga gcaagggatt 2401 caccctccgc aggctcagag caagatagag gagctggagc aagaaccgga gccggagccg 2461 gagccggaac cggagcccga gcccgagccc gagccggagc agctcctcga ggaacaaaaa 2521 ctaatatcag aggaagattt ggaacagaaa ctaatcagtg aagaagattt agagcaaaag 2581 ttaatttccg aggaggactt atgagggccc gtttaaaccc gctgatcagc ctcgactgtg 2641 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 2701 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 2761 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 2821 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 2881 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 2941 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3001 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3061 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3121 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 3181 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 3241 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 3301 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 3361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3421 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3481 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 3541 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 3601 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3661 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 3721 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 3781 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 3841 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 3901 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 3961 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 4021 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 4081 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 4141 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 4201 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 4261 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 4321 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 4381 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 4441 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 4501 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 4561 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 4621 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 4681 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 4741 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 4801 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 4861 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 4921 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 4981 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 5041 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 5101 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 5161 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 5221 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 5281 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 5341 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 5401 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 5461 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 5521 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 5581 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 5641 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 5701 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 5761 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 6361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 6421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 6481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 6541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 6601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 6661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 6721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 6781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 6841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 6901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 6961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7021 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt //