LOCUS Exported 5016 bp ds-DNA circular SYN 03-JAN-2023 DEFINITION Expresses EGFP-FKBP12 fusion protein. ACCESSION . VERSION . KEYWORDS pEGFPC1 FKBP12 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5016) TITLE Jun Liu Lab Plasmids 2022 REFERENCE 2 (bases 1 to 5016) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 3, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5016 /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 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 613..1329 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" CDS 1351..1674 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL LKLE" polyA_signal 1804..1925 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1841..1860) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1895..1914 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1932..2387) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2069..2090) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2281..2300 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2414..2518 /gene="bla" /label=AmpR promoter promoter 2520..2877 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2543..2563) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2728..2863 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2790..2809 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2912..3706 /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(2966..2985) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3576..3595 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3894..3913) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3938..3985 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4314..4902 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4803..4822 /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctcgagct atgggagtgc aggtggaaac catctcccca 1381 ggagacgggc gcaccttccc caagcgcggc cagacctgcg tggtgcacta caccgggatg 1441 cttgaagatg gaaagaaatt tgattcctcc cgggacagaa acaagccctt taagtttatg 1501 ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt 1561 cagagagcca aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc 1621 atcatcccac cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaaggatcc 1681 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct 1741 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt 1801 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 1861 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 1921 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt 2401 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 2461 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc 2521 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2581 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 2641 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2701 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 2761 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 2821 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat 2881 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 2941 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 3001 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 3061 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct 3121 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 3181 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 3241 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 3301 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 3361 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 3421 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 3481 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 3541 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 3601 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 3661 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 3721 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 3781 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 3841 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca 3901 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa 3961 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 4021 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 4081 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 4141 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa 4201 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 4261 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 4321 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 4381 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 4441 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 4501 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 4561 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 4621 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 4681 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 4741 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 4801 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 4861 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 4921 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 4981 cctgattctg tggataaccg tattaccgcc atgcat //