LOCUS       sequence-456532-        5313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mCherry-Rab9a in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5313)
  AUTHORS   Lee JG, Takahama S, Zhang G, Tomarev SI, Ye Y
  TITLE     Unconventional secretion of misfolded proteins promotes adaptation 
            to proteasome dysfunction in mammalian cells.
  JOURNAL   Nat Cell Biol. 2016 Jul;18(7):765-76. doi: 10.1038/ncb3372. Epub 
            2016 Jun 13.
  PUBMED    27295555
REFERENCE   2  (bases 1 to 5313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ncb3372"; 
            journalName: "Nat Cell Biol"; date: "2016-07"; volume: "18"; issue: 
            "7"; pages: "765-76"
COMMENT     Sequence Label: mCherry-Rab9a
FEATURES             Location/Qualifiers
     source          1..5313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..72)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    890..937
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     962..981
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1169..1963)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1169..1963)
                     /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(1280..1299)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1890..1909
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1998..2355)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2012..2147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2164..2355
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2312..2332
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2357..2461)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2488..2943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2575..2594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2785..2806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2950..3071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2961..2980)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3015..3034
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3852..4559)
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     4396..4414
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      4556..4565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4604..4807)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4633..4653)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4808..5111
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(join(5286..5313,1..561))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
       61 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      121 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      181 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      241 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      301 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      361 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      421 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      481 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      541 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      601 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
      661 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct
      721 gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
      781 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
      841 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
      901 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
      961 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     1021 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     1081 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     1141 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     1201 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     1261 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     1321 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     1381 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     1441 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     1501 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     1561 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     1621 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     1681 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     1741 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     1801 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     1861 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     1921 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     1981 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     2041 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     2101 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     2161 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     2221 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     2281 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     2341 ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt
     2401 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2461 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     2521 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2581 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2641 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2701 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2761 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2821 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2881 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     2941 tttacgcgtt aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3001 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3061 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     3121 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt
     3181 tatctagatc cggtggatcc cgggcccgcg gtaccgtcga ctcaacagca agatgagcta
     3241 ggcttgggct ttcggtgaag attgactgtg tctgtctgaa tcaaatgatc tgacctatcc
     3301 tcggtagcaa gaactcttcg aaccgcttcc tcaaaggctg ctgccacatt tgtggcatct
     3361 tttgcacttg tttcaaaata aggatagtcg ccgttgtccc tgcaccaagc ttgggcttct
     3421 tctgtagaca cctgccgttc gcttatgtca atcttgttac ccagaatcac aaaaggaaag
     3481 ctctcaggct ctttcacatc tgcataatat atgaattctt tcttccagtt acttaagttc
     3541 tggaagcttt gtgaatcatc gacactaaaa gtaagcaggc agcagtcaga acctctgtaa
     3601 aatggtgtcc tcaggcttcg gaatcgctcc tgacctgccg tgtcccaaat ctgcatggta
     3661 acaaaatgtc catccacttc caaatcttta tttaaaaatt ccacacctat tgtatggaag
     3721 agctgggtat caaacttatt agttacatat ctgttcataa gtgaactctt cccaactcca
     3781 ccatctccaa ggagaattac tttaaaaagt gatgattttc ctgccatgcg agctcgagat
     3841 ctgagtccgg acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt
     3901 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggaaa ggatgtccaa cttgatgtcg
     3961 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
     4021 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
     4081 ccgtcctcgg ggtacatccg ctcagtggag gcctcccagc ccatggtacg acactgcatt
     4141 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
     4201 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     4261 cgctcccaat tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     4321 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
     4381 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     4441 ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     4501 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatg
     4561 gtggcgaccg gtagcgctag cggatctgac ggttcactaa accagctctg cttatataga
     4621 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     4681 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     4741 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     4801 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     4861 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     4921 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     4981 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     5041 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     5101 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     5161 attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5221 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5281 gcgtttttcc ataggctccg cccccctgac gag
//
