LOCUS       sequence-338230-        5298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human constitutively active form of Rab9a(Q66L) in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5298)
  AUTHORS   Wu H, Voeltz GK
  TITLE     Reticulon-3 Promotes Endosome Maturation at ER Membrane Contact 
            Sites.
  JOURNAL   Dev Cell. 2021 Jan 11;56(1):52-66.e7. doi: 
            10.1016/j.devcel.2020.12.014.
  PUBMED    33434526
REFERENCE   2  (bases 1 to 5298)
  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: 
            "10.1016/j.devcel.2020.12"; journalName: "Dev Cell"; date: 
            "2021-01-11- 11"; volume: "56"; issue: "1"; pages: "52-66.e7"
COMMENT     Sequence Label: GFP-Rab9aQ66L
FEATURES             Location/Qualifiers
     source          1..5298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1329
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2086..2207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2123..2142)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2177..2196
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2214..2669)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2351..2372)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2563..2582
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2696..2800
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2802..3159
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2802..2993
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2825..2845)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3010..3145
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3194..3988
                     /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             3194..3988
                     /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(3248..3267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3858..3877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4176..4195)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4220..4267
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4596..5184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5085..5104
                     /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 atggcaggaa aatcatcact ttttaaagta
     1381 attctccttg gagatggtgg agttgggaag agttcactta tgaacagata tgtaactaat
     1441 aagtttgata cccagctctt ccatacaata ggtgtggaat ttttaaataa agatttggaa
     1501 gtggatggac attttgttac catgcagatt tgggacacgg caggtctgga gcgattccga
     1561 agcctgagga caccatttta cagaggttct gactgctgcc tgcttacttt tagtgtcgat
     1621 gattcacaaa gcttccagaa cttaagtaac tggaagaaag aattcatata ttatgcagat
     1681 gtgaaagagc ctgagagctt tccttttgtg attctgggta acaagattga cataagcgaa
     1741 cggcaggtgt ctacagaaga agcccaagct tggtgcaggg acaacggcga ctatccttat
     1801 tttgaaacaa gtgcaaaaga tgccacaaat gtggcagcag cctttgagga agcggttcga
     1861 agagttcttg ctaccgagga taggtcagat catttgattc agacagacac agtcaatctt
     1921 caccgaaagc ccaagcctag ctcatcttgc tgttgaggat ccaccggatc tagataactg
     1981 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2041 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2101 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2161 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     2221 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2281 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2341 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2401 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2461 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2521 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2581 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2641 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2701 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2761 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     2821 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2881 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2941 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3001 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3061 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3121 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3181 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3241 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3301 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3361 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3421 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3481 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3541 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3601 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3661 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3721 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3781 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3841 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3901 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3961 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4021 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4081 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4141 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4201 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4261 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4321 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4381 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4441 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4501 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4561 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4621 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4681 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4741 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4801 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4861 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4921 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4981 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5041 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5101 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5161 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5221 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5281 cgtattaccg ccatgcat
//
