LOCUS Exported 5418 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS GFP-rab7 WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5418) AUTHORS Choudhury A, Dominguez M, Puri V, Sharma DK, Narita K, Wheatley CL, Marks DL, Pagano RE TITLE Rab proteins mediate Golgi transport of caveola-internalized glycosphingolipids and correct lipid trafficking in Niemann-Pick C cells. JOURNAL J Clin Invest. 2002 Jun . 109(12):1541-50. PUBMED 12070301 REFERENCE 2 (bases 1 to 5418) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5418 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 68..371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 372..575 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 526..546 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 614..623 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 620..1336 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 620..1336 /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(665..686) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(926..945) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1273..1294 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1337..1402 /label=MCS /note="multiple cloning site" CDS 1406..1432 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1433..1459 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2213..2334 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2250..2269) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2304..2323 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2341..2796) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2478..2499) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2690..2709 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2823..2927 /gene="bla" /label=AmpR promoter promoter 2929..3286 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2952..2972) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3137..3272 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3199..3218 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3321..4115 /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(3375..3394) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3985..4004 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4303..4322) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4347..4394 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4723..5311 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5212..5231 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc 661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg 841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa 961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat 1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga 1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga 1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 1321 ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga 1381 cggtaccgcg ggcccgggat ccatgtaccc atacgatgtt ccggattacg cttacccata 1441 cgatgttccg gattacgctg gtacctctag gaagaaagtg ttgctgaagg ttatcatcct 1501 gggagattct ggagtcggga agacatcact catgaaccag tatgtgaata agaaattcag 1561 caatcagtac aaagccacaa taggagctga ctttctgacc aaggaggtga tggtggatga 1621 caggctagtc acaatgcaga tatgggacac agcaggacag gaacggttcc agtctctcgg 1681 tgtggccttc tacagaggtg cagactgctg cgttctggta tttgatgtga ctgcccccaa 1741 cacattcaaa accctagata gctggagaga tgagtttctc atccaggcca gtccccgaga 1801 tcctgaaaac ttcccatttg ttgtgttggg aaacaagatt gacctcgaaa acagacaagt 1861 ggccacaaag cgggcacagg cctggtgcta cagcaaaaac aacattccct actttgagac 1921 cagtgccaag gaggccatca acgtggagca ggcgttccag acgattgcac ggaatgcact 1981 taagcaggaa acggaggtgg agctgtacaa cgaatttcct gaacctatca aactggacaa 2041 gaatgaccgg gccaaggcct cggcagaaag ctgcagttgc tgaggatcca ccggatctag 2101 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct 2161 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt 2221 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 2281 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt 2341 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat 2401 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 2461 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 2521 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct 2581 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc 2641 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag 2701 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca 2761 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg 2821 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 2881 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa 2941 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 3001 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3061 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3121 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3181 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt 3241 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga 3301 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 3361 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 3421 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 3481 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg 3541 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 3601 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 3661 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 3721 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 3781 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 3841 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 3901 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 3961 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 4021 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4081 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 4141 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4201 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4261 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac 4321 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt 4381 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc 4441 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc 4501 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc 4561 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 4621 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 4681 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 4741 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 4801 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 4861 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 4921 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 4981 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 5041 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 5101 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 5161 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 5221 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 5281 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 5341 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 5401 ggataaccgt attaccgc //