LOCUS V039354 8967 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039354 VERSION V039354 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 955..3780 /label="USP33(NM_015017)" /note="USP33(NM_015017)" /gene="USP33" CDS 3817..4536 /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 4562..4786 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4832..5260 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5274..5603 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5670..6464 /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 6640..6761 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6810..6826) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6834..6850 /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(6858..6888) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6903..6924 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7212..7800) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7971..8831) /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(8832..8936) /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 gctggctagt 901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc caccatgaca 961 ggatcaaatt cacacataac gatattaacc ttaaaggtgt tacctcattt tgaaagtctt 1021 gggaaacagg aaaaaattcc taacaaaatg tcagcttttc gaaatcattg tccacatttg 1081 gattcagttg gtgaaataac aaaagaagat ttgatacaaa aatcccttgg tacttgtcag 1141 gattgtaaag tccaaggacc aaatctttgg gcatgtctgg agaatagatg ttcatatgtt 1201 ggctgtggtg aatcacaagt agatcacagc accatacatt ctcaggagac aaagcattat 1261 ctaactgtga accttaccac tcttcgagta tggtgttatg cttgcagcaa agaagtattt 1321 ttggatagga aattaggaac tcagccttca ttgcctcatg taagacaacc tcaccaaata 1381 caagaaaaca gtgtccagga ttttaaaata cccagtaata caacattaaa aactcctctg 1441 gttgccgtat ttgatgatct ggatatagaa gcggatgaag aagatgaact tagggccaga 1501 ggtcttacag gtttgaaaaa tattggaaat acttgttaca tgaatgcagc tttgcaggct 1561 ctttctaatt gcccaccttt gacacagttt tttcttgatt gtggaggact agctcgaaca 1621 gataagaaac ctgccatttg taaaagttat ctcaaactaa tgacagagct gtggcataaa 1681 agcaggccag gatctgttgt gcctactact ctgtttcaag gaattaaaac tgtaaatcca 1741 acatttcggg ggtattctca gcaggatgct caagaattcc ttcgatgttt aatggatttg 1801 cttcatgaag aattgaaaga gcaagtcatg gaagtagaag aagatccgca aaccataacc 1861 actgaggaga caatggaaga agacaagagc cagtcggatg tagattttca gtcttgtgaa 1921 tcttgtagca acagtgatag agcagaaaat gaaaatggct ctagatgctt ttctgaagat 1981 aataatgaaa caacaatgtt aattcaggat gatgaaaaca attcagaaat gtcaaaggat 2041 tggcaaaaag agaagatgtg caataagatt aataaagtaa attctgaagg cgaatttgat 2101 aaagatagag actctatatc tgaaacagtc gacttaaaca accaggaaac tgtcaaagtg 2161 caaatacaca gcagagcttc agaatatatc actgatgtcc attcgaatga cctgtctaca 2221 ccacagatcc ttccatcaaa tgaaggtgtt aatccacgtt tatcggcaag ccctcctaaa 2281 tcaggcaatt tgtggccagg attggcacca ccacacaaaa aagctcagtc tgcatctcca 2341 aagagaaaaa aacagcacaa gaaatacaga agtgttattt cagacatatt tgatggaaca 2401 atcattagtt cagtgcagtg tctgacttgt gacagggtgt ctgtaaccct cgagaccttt 2461 caagatctgt ccttgccaat tcctggcaag gaagaccttg ctaagctgca ttcatcaagt 2521 catccaactt ctatagtcaa agcaggatca tgtggcgaag catatgctcc acaagggtgg 2581 atagcttttt tcatggaata tgtgaagagg tttgttgtct catgtgtccc tagctggttt 2641 tggggtccag tagtaacctt gcaagattgt cttgctgcct tctttgccag agatgaacta 2701 aaaggtgaca atatgtacag ttgtgaaaaa tgcaaaaagt tgagaaatgg agtgaagttt 2761 tgtaaagtac aaaactttcc tgagattttg tgcatccacc ttaaaagatt cagacatgaa 2821 ctaatgtttt ccaccaaaat cagtacccat gtttcatttc cgctagaagg cttggatctt 2881 cagccatttc ttgctaagga tagtccagct caaattgtga catatgatct tctgtcagtc 2941 atttgccatc atggaactgc aagtagtgga cactatatag cctactgccg aaacaatcta 3001 aataatctct ggtatgaatt tgatgatcag agtgtcactg aagtttcaga atctactgta 3061 caaaatgcag aagcttacgt tcttttctat aggaagagca gcgaagaggc acaaaaagag 3121 aggagaagga tatcaaattt attgaacata atggaaccaa gcctccttca gttttatatt 3181 tctcgacagt ggcttaataa atttaagacc tttgccgaac ctggccctat ttcaaataat 3241 gactttcttt gtattcatgg aggtgttcct ccaagaaaag ctggttatat tgaagacctg 3301 gttttgatgc tgcctcagaa catttgggat aacctatata gcaggtatgg tggaggacca 3361 gctgtcaacc atctgtacat ttgtcatact tgccaaattg aggcggagaa aattgaaaaa 3421 agaagaaaaa ctgaattgga aatttttatt cggcttaaca gagcgttcca aaaagaggac 3481 tctccagcta ctttttattg catcagtatg cagtggttta gagaatggga aagttttgtg 3541 aagggtaaag atggagatcc tccaggtcct attgacaata ctaagattgc agtcactaaa 3601 tgtggtaatg tgatgcttag gcaaggagca gattctggcc agatttctga agaaacatgg 3661 aattttctgc agtctattta tggtggaggg cctgaagtta tcctgcgacc tccggttgtt 3721 catgttgatc cagatatact tcaagcagaa gaaaaaattg aagtagaaac tcggtctttg 3781 ctcgagtcta gagggccctt cgggggtgga ggctctatgg tgagcaaggg cgaggagctg 3841 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 3901 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 3961 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc 4021 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 4081 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 4141 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 4201 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 4261 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 4321 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 4381 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg 4441 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 4501 gggatcactc tcggcatgga cgagctgtac aagtaagttt aaacccgctg atcagcctcg 4561 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 4621 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 4681 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 4741 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 4801 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 4861 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 4921 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 4981 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 5041 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 5101 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 5161 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 5221 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 5281 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 5341 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 5401 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 5461 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 5521 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 5581 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 5641 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 5701 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 5761 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 5821 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 5881 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 5941 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 6001 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 6061 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 6121 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 6181 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 6241 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 6301 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 6361 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 6421 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 6481 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 6541 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 6601 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 6661 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 6721 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 6781 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 6841 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 6901 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 6961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 7021 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 7081 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 7141 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 7201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 7261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 7321 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 7381 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 7441 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 7501 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 7561 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 7621 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 7681 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 7741 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 7801 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 7861 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 7921 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 7981 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 8041 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 8101 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 8161 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 8221 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 8281 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 8341 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 8401 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 8461 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 8521 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 8581 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 8641 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 8701 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 8761 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 8821 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 8881 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 8941 tttccccgaa aagtgccacc tgacgtc //