LOCUS V024702 8236 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024702 VERSION V024702 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 907..3042 /label="USP44(NM_032147)" /note="USP44(NM_032147)" /gene="USP44" CDS 3079..3798 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" polyA_signal 3836..4060 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4106..4534 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4548..4877 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4944..5738 /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 5912..6033 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6082..6098) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6106..6122 /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(6130..6160) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6175..6196 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6484..7069) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7240..8100) /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(8101..8205) /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 gccaccatgc tagcaatgga tacgtgcaaa catgttgggc agctgcagct tgctcaagac 961 cattccagcc tcaaccctca gaaatggcac tgtgtggact gcaacacgac cgagtccatt 1021 tgggcttgcc ttagctgctc ccatgttgcc tgtggaagat atattgaaga gcatgcactc 1081 aagcactttc aagaaagcag tcatcctgtt gcattggagg tgaatgagat gtacgttttt 1141 tgttaccttt gtgatgatta tgttctgaat gataacacaa ctggagacct gaagttacta 1201 cgacgtacat taagtgccat caaaagtcaa aattatcact gcacaactcg tagtgggagg 1261 tttttacggt ccatgggtac aggtgatgat tcttatttct tacatgacgg tgcccaatct 1321 ctgcttcaaa gtgaagatca actgtatact gctctttggc acaggagaag gatactaatg 1381 ggtaaaatct ttcgaacatg gtttgaacaa tcacccattg gaagaaaaaa gcaagaagaa 1441 ccatttcagg aaaaaatagt agtaaaaaga gaagtaaaga aaagacggca ggaattggag 1501 tatcaagtta aagcagaatt ggaaagtatg cctccaagaa agagtttacg tttacaaggg 1561 ctcgctcagt cgaccataat agaaatagtt tctgttcagg tgccagcaca aacgccagca 1621 tcaccagcaa aagataaagt actctctacc tcagaaaatg aaatatctca aaaagtcagt 1681 gactcctcag ttaaacgaag gccaatagta actcctggtg taacaggatt gagaaatttg 1741 ggaaatactt gctatatgaa ttctgttctt caggtgttga gtcatttact tatttttcga 1801 caatgttttt taaagcttga tctgaaccaa tggctggcta tgactgctag cgagaagaca 1861 agatcttgta agcatccacc agtcacagat acagtagtat atcaaatgaa tgaatgtcag 1921 gaaaaagata caggttttgt ttgctccaga caatcaagtc tgtcatcagg actaagtggt 1981 ggagcatcaa aaggtagaaa gatggaactt attcagccaa aggagccaac ttcacagtac 2041 atttctcttt gtcatgaatt gcatactttg ttccaagtca tgtggtctgg aaagtgggcg 2101 ttggtctcac catttgctat gctacactca gtgtggagac tcattcctgc ctttcgtggt 2161 tacgcccaac aagacgctca ggaatttctt tgtgaacttt tagataaaat acaacgtgaa 2221 ttagagacaa ctggtaccag tttaccagct cttatcccca cttctcaaag gaaactcatc 2281 aaacaagttc tgaatgttgt aaataacatt tttcatggac aacttcttag tcaggttaca 2341 tgtcttgcat gtgacaacaa atcaaatacc atagaacctt tctgggactt gtcattggag 2401 tttccagaaa ggtatcaatg cagtggaaaa gatattgctt cccagccatg tctggttact 2461 gaaatgttgg ccaaatttac agaaactgaa gctttagaag gaaaaatcta cgtatgtgac 2521 cagtgtaact caaagcgtag aaggttttcc tccaaaccag ttgtactcac agaagcccag 2581 aaacaactta tgatatgcca cctacctcag gttctcagac tgcacctcaa acgattcagg 2641 tggtcaggac gtaataaccg agagaagatt ggtgttcatg ttggctttga ggaaatctta 2701 aacatggagc cctattgctg cagggagacc ctgaaatccc tcagaccaga atgctttatc 2761 tatgacttgt ccgcggtggt gatgcaccat gggaaaggat ttggctcagg gcactacact 2821 gcctactgct ataattctga aggagggttc tgggtacact gcaatgattc caaactaagc 2881 atgtgcacta tggatgaagt atgcaaggct caagcttata tcttgtttta tacccaacga 2941 gttactgaga atggacattc taaacttttg cctccagagc tcctgttggg gagccaacat 3001 cccaatgaag acgctgatac ctcgtctaat gaaatcctta gcggatccgg cggtggagga 3061 agcggcggag gtggatctat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 3121 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 3181 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg 3241 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc 3301 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 3361 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 3421 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 3481 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg 3541 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac 3601 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 3661 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag 3721 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg 3781 gacgagctgt acaagtagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg 3841 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 3901 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 3961 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 4021 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 4081 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 4141 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 4201 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 4261 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 4321 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 4381 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 4441 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 4501 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 4561 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4621 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4681 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 4741 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 4801 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 4861 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 4921 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 4981 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 5041 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 5101 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 5161 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 5221 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 5281 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 5341 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 5401 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 5461 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 5521 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 5581 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 5641 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 5701 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 5761 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 5821 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 5881 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 5941 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6001 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 6061 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6121 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6181 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6241 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6301 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6361 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6421 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6481 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6541 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6601 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6661 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6721 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6781 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6841 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6901 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 6961 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7141 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7201 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7261 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7321 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7381 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7441 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7501 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7561 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7621 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7681 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7741 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7801 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7861 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7921 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7981 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8041 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8101 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8161 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8221 agtgccacct gacgtc //