LOCUS V020001 5764 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020001 VERSION V020001 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 61..364 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 365..568 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 624..1676 /label="ATF4(NM_182810)" /note="ATF4(NM_182810)" /gene="ATF4" CDS 1710..2429 /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 2552..2673 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(2680..3135) /direction=LEFT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3162..3266 /gene="bla" /label="AmpR promoter" promoter 3268..3625 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3660..4454 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4686..4733 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5062..5650 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgagcc accatgaccg aaatgagctt cctgagcagc gaggtgttgg 661 tgggggactt gatgtccccc ttcgaccagt cgggtttggg ggctgaagaa agcctaggtc 721 tcttagatga ttacctggag gtggccaagc acttcaaacc tcatgggttc tccagcgaca 781 aggctaaggc gggctcctcc gaatggctgg ctgtggatgg gttggtcagt ccctccaaca 841 acagcaagga ggatgccttc tccgggacag attggatgtt ggagaaaatg gatttgaagg 901 agttcgactt ggatgccctg ttgggtatag atgacctgga aaccatgcca gatgaccttc 961 tgaccacgtt ggatgacact tgtgatctct ttgcccccct agtccaggag actaataagc 1021 agccccccca gacggtgaac ccaattggcc atctcccaga aagtttaaca aaacccgacc 1081 aggttgcccc cttcaccttc ttacaacctc ttcccctttc cccaggggtc ctgtcctcca 1141 ctccagatca ttcctttagt ttagagctgg gcagtgaagt ggatatcact gaaggagata 1201 ggaagccaga ctacactgct tacgttgcca tgatccctca gtgcataaag gaggaagaca 1261 ccccttcaga taatgatagt ggcatctgta tgagcccaga gtcctatctg gggtctcctc 1321 agcacagccc ctctaccagg ggctctccaa ataggagcct cccatctcca ggtgttctct 1381 gtgggtctgc ccgtcccaaa ccttacgatc ctcctggaga gaagatggta gcagcaaaag 1441 taaagggtga gaaactggat aagaagctga aaaaaatgga gcaaaacaag acagcagcca 1501 ctaggtaccg ccagaagaag agggcggagc aggaggctct tactggtgag tgcaaagagc 1561 tggaaaagaa gaacgaggct ctaaaagaga gggcggattc cctggccaag gagatccagt 1621 acctgaaaga tttgatagaa gaggtccgca aggcaagggg gaagaaaagg gtccccggag 1681 gagggggaag cgatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg 1741 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 1801 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 1861 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt 1921 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 1981 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 2041 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 2101 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 2161 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 2221 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 2281 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 2341 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 2401 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca 2461 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct 2521 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta 2581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 2641 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt 2701 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa 2761 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag 2821 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg 2881 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga 2941 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg 3001 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg 3061 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc 3121 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 3181 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 3241 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg 3301 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 3361 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3421 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 3481 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 3541 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 3601 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca 3661 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 3721 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 3781 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 3841 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 3901 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 3961 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 4021 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 4081 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 4141 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg 4201 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 4261 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 4321 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4381 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4441 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct 4501 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 4561 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 4621 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc 4681 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg 4741 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca 4801 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg 4861 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact 4921 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 4981 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 5041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 5101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 5161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta 5221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 5281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 5341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 5401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 5461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 5521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 5581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 5641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 5701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat 5761 gcat //