LOCUS V038423 8121 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038423 VERSION V038423 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 744..1451 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 1500..2801 /label="Eno2(NM_013509)" /note="Eno2(NM_013509)" /gene="Eno2" CDS 2808..2873 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2897..3485 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3551..3599 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3801..4229 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4243..4572 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4639..5433 /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 5609..5730 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5779..5795) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5803..5819 /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(5827..5857) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5872..5893 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6181..6769) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6940..7800) /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(7801..7905) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca 781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg 841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg 901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg 961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc 1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga 1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca 1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg 1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga 1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc 1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact 1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg 1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca 1501 tgtctataga gaagatttgg gcccgagaga tcttggactc ccgtgggaat cccaccgtgg 1561 aggtggatct ctatactgcc aaaggtcttt tccgggctgc agtccccagt ggagcctcca 1621 ccggcatcta tgaggccctg gaactaaggg atggggacaa acagcgttac ttaggcaaag 1681 gtgtcctgaa ggcagtggac cacatcaaca gcaggattgc accagccctc atcagctcag 1741 gtatctccgt ggtggagcag gagaaactgg acaacctgat gttggaactg gatgggactg 1801 agaataaatc caagtttggg gccaatgcca tcctgggtgt gtccctggcc gtgtgtaagg 1861 ctggggcagc tgagagggac ttgcccctct atcgccacat tgctcagcta gctgggaact 1921 ccgacctcat cctgcctgtg ccggccttta atgtgatcaa tggtggctct catgctggga 1981 acaagttggc catgcaggag ttcatgatcc tcccagtggg tgctgagagc tttcgggatg 2041 ccatgcgact tggggcagag gtgtaccaca ccctcaaggg ggtcatcaag gacaagtatg 2101 gcaaggatgc cactaacgtg ggggatgaag gcggctttgc ccccaatatc ctggagaaca 2161 gcgaagcctt ggagctggtg aaggaagcca tcgacaaggc tggctacacg gaaaagatgg 2221 tgatcggtat ggatgtggct gcctctgagt tttaccgcga tggcaaatac gacttggatt 2281 tcaagtctcc cgctgatcct tcccgataca tcactgggga ccagctgggg gcactctacc 2341 aggactttgt ccggaactat cctgtggtct ccattgaaga cccatttgac caggacgact 2401 gggcagcttg gtccaagttc acagccaacg tcggcatcca gatagtgggc gatgacctga 2461 cggtgaccaa ccccaagcgc atcgagcggg cagtggagga gaaggcctgc aactgtttgc 2521 tgctcaaggt caatcagatc ggctcggtca cagaagccat ccaagcgtgc aagctggccc 2581 aggagaatgg ctggggggtt atggtgagcc atcgctctgg agaaacggag gacacgttca 2641 ttgctgatct tgtcgtcgga ctgtgtacag gccagatcaa gactggcgcc ccatgcagat 2701 ctgaacgtct ggcgaagtac aaccagctca tgagaattga ggaagagctg ggggacgaag 2761 ctcgcttcgc gggacataat ttccgaaatc ccagtgtgct gctcgaggac tacaaagacc 2821 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag 2881 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 2941 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 3001 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 3061 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 3121 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 3181 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 3241 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc 3301 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta 3361 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 3421 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 3481 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc 3541 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca 3601 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg 3661 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg 3721 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg 3781 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 3841 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 3901 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat 3961 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 4021 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 4081 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 4141 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga 4201 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 4321 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 4381 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 4441 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 4501 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 4561 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca 4621 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 4681 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 4741 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 4801 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 4861 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 4921 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 4981 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 5041 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 5101 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 5161 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 5221 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 5281 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 5341 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 5401 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac 5461 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 5521 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 5581 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 5641 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 5701 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta 5761 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5821 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 5881 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5941 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6001 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6061 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6121 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6181 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6301 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6361 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6421 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6481 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6541 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6601 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 6661 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6721 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6781 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6841 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6901 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6961 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7021 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7081 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7141 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7201 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7261 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7321 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7381 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7441 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7501 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7561 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7621 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7681 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7741 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7801 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7861 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7921 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt 7981 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag 8041 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat 8101 tgcatgaaga atctgcttag g //