LOCUS Exported 5253 bp ds-DNA circular SYN 20-APR-2022 DEFINITION Fluorescently tagged HP1alpha. ACCESSION . VERSION . KEYWORDS pGFP-mHP1alpha SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5253) TITLE Rippe Lab Plasmids 2022 REFERENCE 2 (bases 1 to 5253) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 20, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5253 /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" polyA_signal 2041..2162 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2078..2097) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2132..2151 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2169..2624) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2306..2327) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2518..2537 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2651..2755 /gene="bla" /label=AmpR promoter promoter 2757..3114 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2780..2800) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2965..3100 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3027..3046 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3149..3943 /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(3203..3222) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3813..3832 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4131..4150) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4175..4222 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4551..5139 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5040..5059 /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 ccggaatggg aaagaagacc aagaggacag ccgacagctc ttcttcagag 1381 gatgaggagg aatatgtggt ggaaaaggtg ttggacaggc gcatggttaa ggggcaagtg 1441 gaatatctgt tgaagtggaa aggcttttct gaggagcaca atacttggga acctgagaag 1501 aacttggatt gtcctgaact aatttctgag tttatgaaaa agtataagaa gatgaaggag 1561 ggtgaaaaca ataagcccag ggagaaatca gaaggaaaca agaggaaatc cagtttctcc 1621 aacagcgctg atgatattaa atctaaaaaa aagagagagc aaagcaatga tatcgctcgg 1681 ggctttgaga gaggactgga accagaaaag atcatcggag caacagattc ctgcggtgac 1741 ttaatgttct taatgaaatg gaaagacaca gatgaagctg acctggttct tgcaaaagaa 1801 gctaacgtga agtgtccaca gattgtgata gcattttatg aagagagact gacgtggcac 1861 gcatatccag aggatgcgga aaacaaagaa aaagaaagcg cgaagagcta aggatccacc 1921 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta 1981 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt 2041 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 2101 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 2161 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 2221 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 2281 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 2341 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 2401 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 2461 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 2521 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 2581 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg 2641 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 2701 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga 2761 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 2821 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 2881 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2941 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3001 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 3061 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga 3121 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 3181 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 3241 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 3301 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc 3361 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 3421 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 3481 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 3541 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 3601 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 3661 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 3721 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 3781 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 3841 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 3901 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 3961 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 4021 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 4081 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg 4141 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg 4201 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 4261 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 4321 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 4381 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt 4441 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 4501 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 4561 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 4621 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 4681 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 4741 gtatcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 4801 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 4861 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 4921 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 4981 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 5041 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 5101 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 5161 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 5221 gattctgtgg ataaccgtat taccgccatg cat //