LOCUS Exported 5394 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express TagRFP labeled human Rab5a (GTPase located on recycling endosomal membranes).. ACCESSION . VERSION . KEYWORDS pTag-RFP-C-h-Rab4a-c-Myc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5394) AUTHORS Boothe T, Lim GE, Cen H, Skovso S, Piske M, Li SN, Nabi IR, Gilon P, Johnson JD TITLE Inter-domain tagging implicates caveolin-1 in insulin receptor trafficking and Erk signaling bias in pancreatic beta-cells. JOURNAL Mol Metab. 2016 Feb 10;5(5):366-78. doi: 10.1016/j.molmet.2016.01.009. eCollection 2016 May. PUBMED 27110488 REFERENCE 2 (bases 1 to 5394) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5394 /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..1323 /codon_start=1 /product="monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)" /label=TagRFP /note="mammalian codon-optimized" /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV ARYCDLPSKLGHKLN" CDS 2020..2049 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" polyA_signal 2182..2303 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2219..2238) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2273..2292 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2310..2765) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2447..2468) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2659..2678 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2792..2896 /gene="bla" /label=AmpR promoter promoter 2898..3255 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2921..2941) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3106..3241 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3168..3187 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3290..4084 /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(3344..3363) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3954..3973 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4272..4291) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4316..4363 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4692..5280 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5181..5200 /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 ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag 661 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc 721 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc 781 ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac 841 acccagggca tccccgactt ctttaagcag tccttccctg agggcttcac atgggagaga 901 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac 961 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg 1021 atgcagaaga aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc 1081 ggcctggaag gcagaagcga catggccctg aagctcgtgg gcgggggcca cctgatctgc 1141 aacttcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc 1201 tactatgtgg accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag 1261 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt 1321 aattccggac tcagatctcg agctcaaatg tcgcagacgg ccatgtccga aacctacgat 1381 tttttgttta agttcttggt tattggaaat gcaggaactg gcaaatcttg cttacttcat 1441 cagtttattg aaaaaaaatt caaagatgac tcaaatcata caataggagt ggaatttggt 1501 tcaaagataa taaatgttgg tggtaaatat gtaaagttac aaatatggga tacagcagga 1561 caagaacgat tcaggtccgt gacgagaagt tattaccgag gcgcggccgg ggctctcctc 1621 gtctatgata tcaccagccg agaaacctac aatgcgctta ctaattggtt aacagatgcc 1681 cgaatgctag cgagccagaa cattgtgatc atcctttgtg gaaacaagaa ggacctggat 1741 gcagatcgtg aagttacctt cttagaagcc tccagatttg ctcaagaaaa tgagctgatg 1801 tttttggaaa caagtgcgct cacaggggag aatgtagaag aggcttttgt acagtgtgca 1861 agaaaaatac ttaacaaaat cgaatcaggt gagctggacc cagaaagaat gggctcaggt 1921 attcagtacg gagatgctgc cttgagacag ctgaggtcac cgcggcgcgc acaggccccg 1981 aacgctcagg agtgtggttg tacgcgtacg cggccgctcg agcagaaact catctcagaa 2041 gaggatctgt aaggatccac cggatctaga taactgatca taatcagcca taccacattt 2101 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 2161 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 2221 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 2281 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt 2341 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 2401 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 2461 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 2521 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 2581 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 2641 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 2701 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 2761 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 2821 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 2881 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg 2941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3001 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3061 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 3121 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 3181 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 3241 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca 3301 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 3361 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 3421 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc 3481 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 3541 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 3601 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 3661 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 3721 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 3781 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct 3841 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 3901 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 3961 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4021 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4081 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 4141 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 4201 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg 4261 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 4321 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg 4381 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 4441 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 4501 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat 4561 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 4621 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 4681 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 4741 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 4801 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 4861 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 4921 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 4981 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 5041 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 5101 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 5161 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 5221 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 5281 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 5341 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat //