LOCUS Exported 5416 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of GFP-tagged human Rab27A in mammalian cells. ACCESSION . VERSION . KEYWORDS GFP-Rab27A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5416) AUTHORS Westbroek W, Tuchman M, Tinloy B, De Wever O, Vilboux T, Hertz JM, Hasle H, Heilmann C, Helip-Wooley A, Kleta R, Gahl WA TITLE A novel missense mutation (G43S) in the switch I region of Rab27A causing Griscelli syndrome. JOURNAL Mol Genet Metab. 2008 Jun;94(2):248-54. doi: 10.1016/j.ymgme.2008.02.009. Epub 2008 Apr 7. PUBMED 18397837 REFERENCE 2 (bases 1 to 5416) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5416 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 15..603 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 504..523 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 778..1081 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1082..1285 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1236..1256 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1324..1333 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1330..2046 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1330..2046 /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(1375..1396) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1636..1655) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1983..2004 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2921..3042 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2958..2977) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3012..3031 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3049..3504) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3186..3207) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3398..3417 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3531..3635 /gene="bla" /label=AmpR promoter promoter 3637..3994 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3660..3680) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3845..3980 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3907..3926 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4029..4823 /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(4083..4102) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4693..4712 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5011..5030) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5055..5102 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag 721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt 781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac 841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg 901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag 961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat 1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat 1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt 1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga 1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg 1261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg 1321 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc 1381 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat 1441 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc 1501 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac 1561 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc 1621 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc 1681 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc 1741 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag 1801 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg 1861 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc 1921 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat 1981 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg 2041 tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctatgtctga tggagattat 2101 gattacctca tcaagttttt agctttggga gactctggtg tagggaagac cagtgtactt 2161 taccaatata cagatggtaa atttaactcc aaatttatca caacagtggg cattgatttc 2221 agggaaaaaa gagtggtgta cagagccagt gggccggatg gagccactgg cagaggccag 2281 agaatccacc tgcagttatg ggacacagca gggcaggaga ggtttcgtag cttaacgaca 2341 gcgttcttca gagatgctat gggttttctt ctactttttg atctgacaaa tgagcaaagt 2401 ttcctcaatg tcagaaactg gataagccag ctacagatgc atgcatattg tgaaaaccca 2461 gatatagtgc tgtgtggaaa caagagtgat ctggaggacc agagagtagt gaaagaggag 2521 gaagccatag cactcgcaga gaaatatgga atcccctact ttgaaactag tgctgccaat 2581 gggacaaaca taagccaagc aattgagatg cttctggacc tgataatgaa gcgaatggaa 2641 cggtgtgtgg acaagtcctg gattcctgaa ggagtggtgc gatcaaatgg tcatgcctct 2701 acggatcagt taagtgaaga aaaggagaaa ggggcatgtg gctgttgaga agtcaagtaa 2761 gcgacatagt agttcaggtg gcccatgcct gggatccacc ggatctagat aactgatcat 2821 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc 2881 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 2941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 3001 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg 3061 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 3121 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 3181 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 3241 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 3301 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 3361 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 3421 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 3481 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 3541 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 3601 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg 3661 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 3721 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 3841 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 3901 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 3961 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat 4021 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 4081 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 4141 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 4201 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 4261 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 4321 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 4381 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 4441 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 4501 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca 4561 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 4621 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4681 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 4741 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4801 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 4861 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4921 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4981 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg 5041 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt 5101 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca 5161 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga 5221 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc 5281 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 5341 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 5401 agaccccgta gaaaag //