LOCUS Exported 6215 bp ds-DNA circular SYN 20-APR-2022 DEFINITION Fluorescently tagged MECP2, GFP. ACCESSION . VERSION . KEYWORDS pMECP2-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6215) TITLE Rippe Lab Plasmids 2022 REFERENCE 2 (bases 1 to 6215) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 20, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6215 /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" CDS 1783..1803 /codon_start=1 /product="6xHis affinity tag" /label=7xHis /translation="HHHHHHH" regulatory 2155..2164 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2161..2880 /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 2161..2877 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2206..2227) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2467..2486) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2814..2835 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3003..3124 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3040..3059) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3094..3113 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3131..3586) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3268..3289) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3480..3499 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3613..3717 /gene="bla" /label=AmpR promoter promoter 3719..4076 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3742..3762) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3927..4062 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3989..4008 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4111..4905 /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(4165..4184) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4775..4794 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5093..5112) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5137..5184 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5513..6101 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6002..6021 /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 attcctatgg ccgccgctgc cgccaccgcc 661 gccgccgccg ccgcgccgag cggaggagga ggaggaggcg aggaggagag actggaggaa 721 aagtcagaag accaggatct ccagggcctc agagacaagc cactgaagtt taagaaggcg 781 aagaaagaca agaaggagga caaagaaggc aagcatgagc cactacaacc ttcagcccac 841 cattctgcag agccagcaga ggcaggcaaa gcagaaacat cagaaagctc aggctctgcc 901 ccagcagtgc cagaagcctc ggcttccccc aaacagcggc gctccattat ccgtgaccgg 961 ggacctatgt atgatgaccc caccttgcct gaaggttgga cacgaaagct taaacaaagg 1021 aagtctggcc gatctgctgg aaagtatgat gtatatttga tcaatcccca gggaaaagct 1081 tttcgctcta aagtagaatt gattgcatac tttgaaaagg tgggagacac ctccttggac 1141 cctaatgatt ttgacttcac ggtaactggg agagggagcc cctccaggag agagcagaaa 1201 ccacctaaga agcccaaatc tcccaaagct ccaggaactg gcaggggtcg gggacgcccc 1261 aaagggagcg gcactgggag accaaaggca gcagcatcag aaggtgttca ggtgaaaagg 1321 gtcctggaga agagccctgg gaaacttgtt gtcaagatgc ctttccaagc atcgcctggg 1381 ggtaagggtg agggaggtgg ggctaccaca tctgcccagg tcatggtgat caaacgccct 1441 ggcagaaagc gaaaagctga agctgacccc caggccattc ctaagaaacg gggtagaaag 1501 cctgggagtg tggtggcagc tgctgcagct gaggccaaaa agaaagccgt gaaggagtct 1561 tccatacggt ctgtgcatga gactgtgctc cccatcaaga agcgcaagac ccgggagacg 1621 gtcagcatcg aggtcaagga agtggtgaag cccctgctgg tgtccaccct tggtgagaaa 1681 agcgggaagg gactgaagac ctgcaagagc cctgggcgta aaagcaagga gagcagcccc 1741 aaggggcgca gcagcagtgc ctcctcccca cctaagaagg agcaccatca tcaccaccat 1801 cactcagagt ccacaaaggc ccccatgcca ctgctcccat ccccaccccc acctgagcct 1861 gagagctctg aggaccccat cagcccccct gagcctcagg acttgagcag cagcatctgc 1921 aaagaagaga agatgccccg aggaggctca ctggaaagcg atggctgccc caaggagcca 1981 gctaagactc agcctatggt cgccaccact accacagttg cagaaaagta caaacaccga 2041 ggggagggag agcgcaaaga cattgtttca tcttccatgc caaggccaaa cagagaggag 2101 cctgtggaca gccggacgcc cgtgaccgag agagttagcg gggatccacc ggtcgccacc 2161 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 2221 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 2281 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 2341 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 2401 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 2461 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 2521 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 2581 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 2641 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 2701 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 2761 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 2821 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa 2881 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt 2941 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg 3001 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 3061 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 3121 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa 3181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa 3241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac 3301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa 3361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct 3421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa 3481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc 3541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt 3601 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 3661 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct 3721 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3901 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 3961 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 4021 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc 4081 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt 4141 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 4201 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 4261 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg 4321 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 4381 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 4441 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 4501 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 4561 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 4621 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 4681 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 4741 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 4801 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 4861 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt 4921 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg 4981 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 5041 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac 5101 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 5161 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg 5221 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac 5281 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc 5341 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat 5401 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 5461 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 5521 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 5581 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 5641 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 5701 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 5761 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 5821 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 5881 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 5941 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 6001 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 6061 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 6121 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 6181 ctgattctgt ggataaccgt attaccgcca tgcat //