LOCUS       sequence-254615-        8280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human Rtn4a fluorescently tagged with GFP on the C-terminus for 
            mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8280)
  AUTHORS   Shibata Y, Voss C, Rist JM, Hu J, Rapoport TA, Prinz WA, Voeltz GK
  TITLE     The reticulon and DP1/Yop1p proteins form immobile oligomers in the 
            tubular endoplasmic reticulum.
  JOURNAL   J Biol Chem. 2008 Jul 4. 283(27):18892-904.
  PUBMED    18442980
REFERENCE   2  (bases 1 to 8280)
  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"; journalName: "J Biol 
            Chem. 2008 Jul 4. 283(27):18892-904."
COMMENT     Sequence Label: Rtn4a-GFP
FEATURES             Location/Qualifiers
     source          1..8280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4167..4886
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    5008..5129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5045..5064)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5099..5118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5136..5591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5273..5294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5485..5504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5618..5722
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5724..6081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5724..5915
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5747..5767)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5932..6067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6116..6910
                     /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             6116..6910
                     /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(6170..6189)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6780..6799
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7098..7117)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7142..7189
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7518..8106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8007..8026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct atggaagacc tggaccagtc tcctctggtc tcgtcctcgg
      601 acagcccacc ccggccgcag cccgcgttca agtaccagtt cgtgagggag cccgaggacg
      661 aggaggaaga agaggaggag gaagaggagg acgaggacga agacctggag gagctggagg
      721 tgctggagag gaagcccgcc gccgggctgt ccgcggcccc agtgcccacc gcccctgccg
      781 ccggcgcgcc cctgatggac ttcggaaatg acttcgtgcc gccggcgccc cggggacccc
      841 tgccggccgc tccccccgtc gccccggagc ggcagccgtc ttgggacccg agcccggtgt
      901 cgtcgaccgt gcccgcgcca tccccgctgt ctgctgccgc agtctcgccc tccaagctcc
      961 ctgaggacga cgagcctccg gcccggcctc cccctcctcc cccggccagc gtgagccccc
     1021 aggcagagcc cgtgtggacc ccgccagccc cggctcccgc cgcgcccccc tccaccccgg
     1081 ccgcgcccaa gcgcaggggc tcctcgggct cagtggatga gacccttttt gctcttcctg
     1141 ctgcatctga gcctgtgata cgctcctctg cagaaaatat ggacttgaag gagcagccag
     1201 gtaacactat ttcggctggt caagaggatt tcccatctgt cctgcttgaa actgctgctt
     1261 ctcttccttc tctgtctcct ctctcagccg cttctttcaa agaacatgaa taccttggta
     1321 atttgtcaac agtattaccc actgaaggaa cacttcaaga aaatgtcagt gaagcttcta
     1381 aagaggtctc agagaaggca aaaactctac tcatagatag agatttaaca gagttttcag
     1441 aattagaata ctcagaaatg ggatcatcgt tcagtgtctc tccaaaagca gaatctgccg
     1501 taatagtagc aaatcctagg gaagaaataa tcgtgaaaaa taaagatgaa gaagagaagt
     1561 tagttagtaa taacatcctt cataatcaac aagagttacc tacagctctt actaaattgg
     1621 ttaaagagga tgaagttgtg tcttcagaaa aagcaaaaga cagttttaat gaaaagagag
     1681 ttgcagtgga agctcctatg agggaggaat atgcagactt caaaccattt gagcgagtat
     1741 gggaagtgaa agatagtaag gaagatagtg atatgttggc tgctggaggt aaaatcgaga
     1801 gcaacttgga aagtaaagtg gataaaaaat gttttgcaga tagccttgag caaactaatc
     1861 acgaaaaaga tagtgagagt agtaatgatg atacttcttt ccccagtacg ccagaaggta
     1921 taaaggatcg ttcaggagca tatatcacat gtgctccctt taacccagca gcaactgaga
     1981 gcattgcaac aaacattttt cctttgttag gagatcctac ttcagaaaat aagaccgatg
     2041 aaaaaaaaat agaagaaaag aaggcccaaa tagtaacaga gaagaatact agcaccaaaa
     2101 catcaaaccc ttttcttgta gcagcacagg attctgagac agattatgtc acaacagata
     2161 atttaacaaa ggtgactgag gaagtcgtgg caaacatgcc tgaaggcctg actccagatt
     2221 tagtacagga agcatgtgaa agtgaattga atgaagttac tggtacaaag attgcttatg
     2281 aaacaaaaat ggacttggtt caaacatcag aagttatgca agagtcactc tatcctgcag
     2341 cacagctttg cccatcattt gaagagtcag aagctactcc ttcaccagtt ttgcctgaca
     2401 ttgttatgga agcaccattg aattctgcag ttcctagtgc tggtgcttcc gtgatacagc
     2461 ccagctcatc accattagaa gcttcttcag ttaattatga aagcataaaa catgagcctg
     2521 aaaacccccc accatatgaa gaggccatga gtgtatcact aaaaaaagta tcaggaataa
     2581 aggaagaaat taaagagcct gaaaatatta atgcagctct tcaagaaaca gaagctcctt
     2641 atatatctat tgcatgtgat ttaattaaag aaacaaagct ttctgctgaa ccagctccgg
     2701 atttctctga ttattcagaa atggcaaaag ttgaacagcc agtgcctgat cattctgagc
     2761 tagttgaaga ttcctcacct gattctgaac cagttgactt atttagtgat gattcaatac
     2821 ctgacgttcc acaaaaacaa gatgaaactg tgatgcttgt gaaagaaagt ctcactgaga
     2881 cttcatttga gtcaatgata gaatatgaaa ataaggaaaa actcagtgct ttgccacctg
     2941 agggaggaaa gccatatttg gaatctttta agctcagttt agataacaca aaagataccc
     3001 tgttacctga tgaagtttca acattgagca aaaaggagaa aattcctttg cagatggagg
     3061 agctcagtac tgcagtttat tcaaatgatg acttatttat ttctaaggaa gcacagataa
     3121 gagaaactga aacgttttca gattcatctc caattgaaat tatagatgag ttccctacat
     3181 tgatcagttc taaaactgat tcattttcta aattagccag ggaatatact gacctagaag
     3241 tatcccacaa aagtgaaatt gctaatgccc cggatggagc tgggtcattg ccttgcacag
     3301 aattgcccca tgacctttct ttgaagaaca tacaacccaa agttgaagag aaaatcagtt
     3361 tctcagatga cttttctaaa aatgggtctg ctacatcaaa ggtgctctta ttgcctccag
     3421 atgtttctgc tttggccact caagcagaga tagagagcat agttaaaccc aaagttcttg
     3481 tgaaagaagc tgagaaaaaa cttccttccg atacagaaaa agaggacaga tcaccatctg
     3541 ctatattttc agcagagctg agtaaaactt cagttgttga cctcctgtac tggagagaca
     3601 ttaagaagac tggagtggtg tttggtgcca gcctattcct gctgctttca ttgacagtat
     3661 tcagcattgt gagcgtaaca gcctacattg ccttggccct gctctctgtg accatcagct
     3721 ttaggatata caagggtgtg atccaagcta tccagaaatc agatgaaggc cacccattca
     3781 gggcatatct ggaatctgaa gttgctatat ctgaggagtt ggttcagaag tacagtaatt
     3841 ctgctcttgg tcatgtgaac tgcacgataa aggaactcag gcgcctcttc ttagttgatg
     3901 atttagttga ttctctgaag tttgcagtgt tgatgtgggt atttacctat gttggtgcct
     3961 tgtttaatgg tctgacacta ctgattttgg ctctcatttc actcttcagt gttcctgtta
     4021 tttatgaacg gcatcaggca cagatagatc attatctagg acttgcaaat aagaatgtta
     4081 aagatgctat ggctaaaatc caagcaaaaa tccctggatt gaagcgcaaa gctgaaacgg
     4141 taccgcgggc ccgggatcca ccggtcatgg tgagcaaggg cgccgagctg ttcaccggca
     4201 tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg ccacaagttc agcgtgagcg
     4261 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     4321 gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct gagctacggc gtgcagtgct
     4381 tctcacgcta ccccgatcac atgaagcagc acgacttctt caagagcgcc atgcctgagg
     4441 gctacatcca ggagcgcacc atcttcttcg aggatgacgg caactacaag tcgcgcgccg
     4501 aggtgaagtt cgagggcgat accctggtga atcgcatcga gctgaccggc accgatttca
     4561 aggaggatgg caacatcctg ggcaataaga tggagtacaa ctacaacgcc cacaatgtgt
     4621 acatcatgac cgacaaggcc aagaatggca tcaaggtgaa cttcaagatc cgccacaaca
     4681 tcgaggatgg cagcgtgcag ctggccgacc actaccagca gaataccccc atcggcgatg
     4741 gccctgtgct gctgcccgat aaccactacc tgtccaccca gagcgccctg tccaaggacc
     4801 ccaacgagaa gcgcgatcac atgatctact tcggcttcgt gaccgccgcc gccatcaccc
     4861 acggcatgga tgagctgtac aagtgagcgg ccgcgactct agatcataat cagccatacc
     4921 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4981 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     5041 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5101 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     5161 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5221 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5281 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5341 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5401 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5461 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5521 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5581 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5641 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5701 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     5761 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5821 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5881 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5941 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6001 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6061 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     6121 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     6181 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     6241 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     6301 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     6361 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     6421 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     6481 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     6541 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     6601 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     6661 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6721 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6781 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6841 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6901 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6961 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     7021 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     7081 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     7141 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     7201 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     7261 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     7321 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     7381 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7501 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7561 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7621 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7681 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7741 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7801 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7861 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7921 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7981 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8041 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8101 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8161 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     8221 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
