LOCUS V021799 6746 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021799 VERSION V021799 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..2236 /label="Fgl2(NM_008013)" /note="Fgl2(NM_008013)" /gene="Fgl2" CDS 2243..2308 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2346..2570 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2616..3044 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3058..3387 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3454..4248 /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 4422..4543 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4592..4608) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4616..4632 /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(4640..4670) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4685..4706 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4994..5579) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5750..6610) /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(6611..6715) /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 atgaggcttc ctggttggtt 961 gtggctgagt tctgccgtcc tcgctgcctg ccgagcggtg gaggagcaca acctgactga 1021 ggggctggag gatgccagcg cccaggctgc ctgccccgcg aggctggagg gcagcgggag 1081 gtgcgagggg agccagtgcc ccttccagct caccctgccc acgctgacca tccagctccc 1141 gcggcagctt ggcagcatgg aggaggtgct caaagaagtg cggaccctca aggaagcagt 1201 ggacagtctg aagaaatcct gccaggactg taagttgcag gctgacgacc atcgagatcc 1261 cggcgggaat ggagggaatg gagcagagac agccgaggac agtagagtcc aggaactgga 1321 gagtcaggtg aacaagctgt cctcagagct gaagaatgca aaggaccaga tccaggggct 1381 gcaggggcgc ctggagacgc tccatctggt aaatatgaac aacattgaga actacgtgga 1441 caacaaagtg gcaaatctaa ccgttgtggt caacagtttg gatggcaagt gttccaagtg 1501 tcccagccaa gaacacatgc agtcacagcc ggttcaacat ctaatataca aagattgttc 1561 cgaccactac gtgctaggaa ggagaagcag tggggcctac agagttaccc ctgatcacag 1621 aaacagcagc tttgaggtct actgtgacat ggagaccatg ggtggaggct ggacggtgct 1681 gcaggctcgc cttgatggca gcaccaactt caccagagag tggaaagact acaaagccgg 1741 ctttggaaac cttgaacgag aattttggtt gggcaacgat aaaattcatc ttctgaccaa 1801 gagtaaggaa atgattttga gaatagatct tgaagacttt aatggtctca cactttatgc 1861 cttgtatgat cagttttatg tggctaatga atttctcaaa taccgattac acatcggtaa 1921 ctacaatggc acggcagggg atgccttgcg tttcagtcga cactacaacc atgacctgag 1981 gtttttcaca accccagaca gagacaacga tcggtacccc tctgggaact gtgggctcta 2041 ttacagctca ggctggtggt ttgattcatg tctctctgcc aacttaaatg gcaaatatta 2101 ccaccagaaa tacaaaggtg tccgtaatgg gattttctgg ggcacctggc ctggtataaa 2161 ccaggcacag ccaggtggct acaagtcctc cttcaaacag gccaagatga tgattaggcc 2221 caagaatttc aagccactcg aggactacaa agaccatgac ggtgattata aagatcatga 2281 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 2341 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 2401 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 2461 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 2521 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 2581 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 2641 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 2701 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 2761 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 2821 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 2881 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 2941 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 3001 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 3061 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 3181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 3241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 3301 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 3361 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 3421 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 3481 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 3541 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 3601 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 3661 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 3721 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 3781 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 3841 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 3901 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 3961 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 4021 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 4081 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 4141 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 4201 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 4261 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 4321 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 4381 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 4441 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 4501 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 4561 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4621 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4681 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 4741 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4801 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4861 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4921 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4981 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5041 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5101 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5161 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 5221 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5281 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5341 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5401 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 5461 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5521 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5581 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5641 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5701 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 5761 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5821 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5881 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 5941 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 6001 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 6061 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 6121 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 6181 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 6241 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 6301 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 6361 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 6421 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 6481 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 6541 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6601 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6661 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 6721 ttccccgaaa agtgccacct gacgtc //