LOCUS Exported 5355 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Human Rab5B fluorescently tagged with GFP on the N-terminus for mammalian expression. ACCESSION . VERSION . KEYWORDS GFP-Rab5B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5355) AUTHORS Rowland AA, Chitwood PJ, Phillips MJ, Voeltz GK TITLE ER Contact Sites Define the Position and Timing of Endosome Fission. JOURNAL Cell. 2014 Nov 20;159(5):1027-41. doi: 10.1016/j.cell.2014.10.023. PUBMED 25416943 REFERENCE 2 (bases 1 to 5355) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5355 /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="Aequorea coerulescens GFP" /label=AcGFP1 /note="mammalian codon-optimized" /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF GFVTAAAITHGMDELYK" polyA_signal 2143..2264 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2180..2199) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2234..2253 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2271..2726) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2408..2429) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2620..2639 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2753..2857 /gene="bla" /label=AmpR promoter promoter 2859..3216 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2882..2902) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3067..3202 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3129..3148 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3251..4045 /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(3305..3324) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3915..3934 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4233..4252) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4277..4324 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4653..5241 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5142..5161 /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 caagggcgcc gagctgttca ccggcatcgt gcccatcctg 661 atcgagctga atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg 781 ccctggccca ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc 841 gatcacatga agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag 901 cgcaccatct tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag 961 ggcgataccc tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac 1021 atcctgggca ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac 1081 aaggccaaga atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc 1141 gtgcagctgg ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg 1201 cccgataacc actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc 1261 gatcacatga tctacttcgg cttcgtgacc gccgccgcca tcacccacgg catggatgag 1321 ctgtacaagt ccggactcag atctcgagct atgactagca gaagcacagc taggcccaat 1381 gggcaacccc aggccagcaa aatttgccag ttcaaattgg tcctgctggg agaatctgca 1441 gtgggaaagt caagcctggt attacgtttt gtcaaagggc agttccatga gtaccaggag 1501 agcaccattg gagcggcctt cctcacccag tccgtttgtc tagatgacac aacagtgaag 1561 tttgagatct gggacacagc tgggcaggag cgatatcaca gcttagcccc catgtactac 1621 aggggtgccc aagctgcaat cgtggtttac gacattacta atcaggaaac ctttgcccga 1681 gcaaagacat gggtgaagga actacagcga caggccagtc ctagcatcgt tattgccctg 1741 gcagggaaca aagctgacct ggccaacaaa cgtatggtgg agtatgaaga ggcccaggca 1801 tatgcagatg acaacagctt attgttcatg gagacttcag ccaagacagc tatgaacgtg 1861 aatgatctct tcctggcaat agctaagaag ttgccaaaga gtgaacccca gaatctggga 1921 ggtgcagcag gccgaagccg gggtgtggat ctccatgaac agtcccagca gaacaagagc 1981 cagtgttgta gcaactgagg taccgcgggc ccgggatcca ccggatctag ataactgatc 2041 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc 2101 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct 2161 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 2221 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag 2281 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca 2341 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag 2401 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg 2461 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt 2521 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag 2581 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc 2641 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc 2701 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac 2761 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 2821 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg 2881 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2941 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3061 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3121 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag 3181 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg 3241 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga 3301 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt 3361 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct 3421 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg 3481 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt 3541 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc 3601 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc 3661 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga 3721 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag 3781 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat 3841 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg 3901 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc 3961 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta 4021 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg 4081 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc 4141 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 4201 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa 4261 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt 4321 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 4381 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 4441 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac 4501 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 4561 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 4621 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 4681 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 4741 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt 4801 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 4861 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 4921 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 4981 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 5041 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 5101 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 5161 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 5221 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 5281 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt 5341 attaccgcca tgcat //