LOCUS       sequence-338309-        5277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human dominant negative form of Rab9a(S21N) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5277)
  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 5277)
  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: mScarlet-Rab9aS21N
FEATURES             Location/Qualifiers
     source          1..5277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             650..1345
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(783..801)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1051..1070
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2102..2223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2139..2158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2193..2212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2257..2685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2367..2388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2579..2598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2712..2816
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2818..3175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2818..3009
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2841..2861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3026..3161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3210..4004
                     /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             3210..4004
                     /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(3264..3283)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3874..3893
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4192..4211)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4236..4283
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4612..5200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5101..5120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa
      661 gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa
      721 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      781 cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc
      841 tcagttcatg tacggctcca gggccttcac caagcacccc gccgacatcc ccgactacta
      901 taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      961 cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa
     1021 gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg
     1081 ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat
     1141 ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc
     1201 caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc
     1261 ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac
     1321 cggcggcatg gacgagctgt acaagtccgg actcagatct cgagctatgg caggaaaatc
     1381 atcacttttt aaagtaattc tccttggaga tggtggagtt gggaagaatt cacttatgaa
     1441 cagatatgta actaataagt ttgataccca gctcttccat acaataggtg tggaattttt
     1501 aaataaagat ttggaagtgg atggacattt tgttaccatg cagatttggg acacggcagg
     1561 tcaggagcga ttccgaagcc tgaggacacc attttacaga ggttctgact gctgcctgct
     1621 tacttttagt gtcgatgatt cacaaagctt ccagaactta agtaactgga agaaagaatt
     1681 catatattat gcagatgtga aagagcctga gagctttcct tttgtgattc tgggtaacaa
     1741 gattgacata agcgaacggc aggtgtctac agaagaagcc caagcttggt gcagggacaa
     1801 cggcgactat ccttattttg aaacaagtgc aaaagatgcc acaaatgtgg cagcagcctt
     1861 tgaggaagcg gttcgaagag ttcttgctac cgaggatagg tcagatcatt tgattcagac
     1921 agacacagtc aatcttcacc gaaagcccaa gcctagctca tcttgctgtt gaggatccac
     1981 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2041 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2101 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2161 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2221 ttaaggcgta aattataagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2281 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2341 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2401 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2461 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2521 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2581 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2641 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     2701 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2761 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     2821 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2881 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2941 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3001 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3061 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3121 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3181 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3241 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3301 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3361 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3421 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3481 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3541 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3601 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3661 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3721 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3781 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3841 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3901 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3961 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4021 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4081 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4141 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4201 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4261 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4321 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4381 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4441 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4501 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4561 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4621 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4681 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4741 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4801 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4861 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4921 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4981 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5041 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5101 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5161 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5221 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatc
//
