LOCUS Exported 5250 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses Arf6Q67L-EGFP in mammalian cells. ACCESSION . VERSION . KEYWORDS Arf6Q67L-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5250) TITLE Scidmore Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 5250) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5250 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1190..1199 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1196..1915 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1196..1912 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1241..1262) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1502..1521) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1849..1870 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2038..2159 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2075..2094) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2129..2148 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2166..2621) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2303..2324) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2515..2534 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2648..2752 /gene="bla" /label=AmpR promoter promoter 2754..3111 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2777..2797) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2962..3097 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3024..3043 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3146..3940 /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" primer_bind complement(3200..3219) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3810..3829 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4128..4147) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4172..4219 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4548..5136 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5037..5056 /label=pBR322ori-F /note="pBR322 origin, forward primer" 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 atctcgagct caagcttcga attcatgggg aaggtgctat ccaaaatctt 661 cgggaacaag gaaatgcgga tcctcatgtt gggcctggac gcggccggca agacaacaat 721 cctgtacaag ttgaagctgg gccagtcggt gaccaccatt cccactgtgg gtttcaacgt 781 ggagacggtg acttacaaaa atgtcaagtt caacgtatgg gatgtgggcg gcctggacaa 841 gatccggccg ctctggcggc attactacac tgggacccaa ggtctcatct tcgtagtgga 901 ctgcgccgac cgcgaccgca tcgatgaggc tcgccaggag ctgcaccgca ttatcaatga 961 ccgggagatg agggacgcca taatcctcat cttcgccaac aagcaggacc tgcccgatgc 1021 catgaaaccc cacgagatcc aggagaaact gggcctgacc cggattcggg acaggaactg 1081 gtatgtgcag ccctcctgtg ccacctcagg ggacggactc tatgaggggc tcacatggtt 1141 aacctctaac tacaaatctg tcgacggtac cgcgggcccg ggatccatcg ccaccatggt 1201 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga 1261 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa 1321 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt 1381 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca 1441 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa 1501 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa 1561 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct 1621 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat 1681 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca 1741 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct 1801 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct 1861 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg 1921 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa 1981 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac 2041 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 2101 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa 2161 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 2221 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 2281 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 2341 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 2401 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 2461 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 2521 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 2581 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg 2641 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 2701 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc 2761 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 2881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 2941 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 3001 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 3061 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca 3121 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 3181 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 3241 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 3301 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac 3361 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 3421 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 3481 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 3541 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 3601 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 3661 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 3721 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 3781 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 3841 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 3901 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa 3961 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 4021 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 4081 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag 4141 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 4201 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 4261 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 4321 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc 4381 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 4441 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 4501 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 4561 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 4621 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 4681 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 4741 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 4801 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 4861 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 4921 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 4981 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 5041 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 5101 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 5161 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 5221 tctgtggata accgtattac cgccatgcat //