LOCUS Exported 5853 bp ds-DNA circular SYN 14-JAN-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5853) AUTHORS . TITLE Direct Submission JOURNAL Exported Tuesday, Jan 26, 2021 from SnapGene Viewer 5.2.4 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5853 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 212..259 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 588..1176 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" enhancer 1351..1654 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1655..1858 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" misc_feature 1925..3073 /label=SAV1 coding regulatory 3083..3092 /regulatory_class="other" /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3089..3808 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" polyA_signal 3931..4052 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(4059..4514) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4541..4645 /gene="bla" /label=AmpR promoter promoter 4647..5004 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4855..4990 /label=SV40 ori /note="SV40 origin of replication" CDS 5039..5833 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" ORIGIN 1 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 61 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 121 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag 181 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 241 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 301 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 361 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc 421 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 481 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 721 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 841 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 901 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 961 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 1021 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 1081 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 1141 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 1201 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 1261 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc 1321 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc 1381 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt 1441 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca 1501 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg 1561 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca 1621 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa 1681 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa 1741 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc 1801 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg 1861 tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct caagcttcga 1921 attcatgctg tcccgaaaga aaaccaaaaa cgaagtgtcc aagccggccg aggtgcaggg 1981 gaagtacgtg aagaaggaga cgtcgcctct gcttcggaat cttatgcctt cattcatccg 2041 gcatggtcca acaattccaa gacgaactga tatctgtctt ccagattcaa gccctaatgc 2101 cttttcaact tctggagatg tagtttcaag aaaccagagt ttccttagaa ctccaattca 2161 aagaacacct catgaaataa tgagaagaga aagcaacaga ttatctgcac cttcttatct 2221 tgccagaagt ctagcagatg tccctagaga gtatggttct tctcagtcat ttgtaacgga 2281 agttagtttt gctgttgaaa atggagactc tggttcccga tattattatt cagacaattt 2341 ttttgatggt cagagaaagc ggccacttgg agatcgtgca catgaagact acagatatta 2401 tgaatacaac catgatctct tccaaagaat gccacagaat caggggaggc atgcttcagg 2461 tattgggaga gttgctgcta catctttagg aaatttgact aaccatggtt ctgaagattt 2521 accccttcct cctggctggt ctgtggactg gacaatgaga gggagaaaat attatataga 2581 tcataacaca aatacaactc actggagcca tcctcttgag cgagaaggac ttcctcctgg 2641 atgggaacga gttgagtcat ccgaatttgg aacctattat gtagatcaca caaataagaa 2701 ggcccaatac aggcatccct gtgctcctag tgtacctcgg tatgatcaac cacctcctgt 2761 cacataccag ccacagcaaa ctgaaagaaa tcagtccctt ctggtacctg caaatccata 2821 tcatactgca gaaattcctg actggcttca ggtttacgca cgagcccctg tgaaatatga 2881 ccacattctg aagtgggaac tcttccagct ggctgacctg gatacatacc agggaatgct 2941 aaagttgctc ttcatgaaag aattggagca gattgttaaa atgtatgaag catacagaca 3001 agcccttctt acagagttgg aaaaccgaaa gcagagacag cagtggtatg cccaacaaca 3061 tggaaaaaat tttccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg 3121 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc 3181 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac 3241 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg 3301 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga 3361 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc 3421 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt 3481 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt 3541 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa 3601 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga 3661 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga 3721 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac 3781 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat 3841 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg 3901 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac 3961 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 4021 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt 4081 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat 4141 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt 4201 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt 4261 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag 4321 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg 4381 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc 4441 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc 4501 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 4561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 4621 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt 4681 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 4741 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 4801 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 4861 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 4921 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 4981 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat 5041 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 5101 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 5161 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 5221 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 5281 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 5341 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 5401 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 5461 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 5521 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg 5581 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 5641 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 5701 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 5761 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 5821 cgagttcttc tgagcgggac tctggggttc gaa //