LOCUS       sequence-338227-        8277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Rtn4a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8277)
  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 8277)
  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: Rtn4a-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..8277
                     /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      4227..4236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4233..4943
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      4488..4497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5065..5186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5102..5121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5156..5175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5193..5648)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5330..5351)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5542..5561
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5675..5779
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5781..6138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5781..5972
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5804..5824)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5989..6124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6173..6967
                     /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             6173..6967
                     /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(6227..6246)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6837..6856
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7155..7174)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7199..7246
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7575..8163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8064..8083
                     /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 ccggactcag atctcgagct atggaagacc tggaccagtc tcctctggtc tcgtcctcgg
      661 acagcccacc ccggccgcag cccgcgttca agtaccagtt cgtgagggag cccgaggacg
      721 aggaggaaga agaggaggag gaagaggagg acgaggacga agacctggag gagctggagg
      781 tgctggagag gaagcccgcc gccgggctgt ccgcggcccc agtgcccacc gcccctgccg
      841 ccggcgcgcc cctgatggac ttcggaaatg acttcgtgcc gccggcgccc cggggacccc
      901 tgccggccgc tccccccgtc gccccggagc ggcagccgtc ttgggacccg agcccggtgt
      961 cgtcgaccgt gcccgcgcca tccccgctgt ctgctgccgc agtctcgccc tccaagctcc
     1021 ctgaggacga cgagcctccg gcccggcctc cccctcctcc cccggccagc gtgagccccc
     1081 aggcagagcc cgtgtggacc ccgccagccc cggctcccgc cgcgcccccc tccaccccgg
     1141 ccgcgcccaa gcgcaggggc tcctcgggct cagtggatga gacccttttt gctcttcctg
     1201 ctgcatctga gcctgtgata cgctcctctg cagaaaatat ggacttgaag gagcagccag
     1261 gtaacactat ttcggctggt caagaggatt tcccatctgt cctgcttgaa actgctgctt
     1321 ctcttccttc tctgtctcct ctctcagccg cttctttcaa agaacatgaa taccttggta
     1381 atttgtcaac agtattaccc actgaaggaa cacttcaaga aaatgtcagt gaagcttcta
     1441 aagaggtctc agagaaggca aaaactctac tcatagatag agatttaaca gagttttcag
     1501 aattagaata ctcagaaatg ggatcatcgt tcagtgtctc tccaaaagca gaatctgccg
     1561 taatagtagc aaatcctagg gaagaaataa tcgtgaaaaa taaagatgaa gaagagaagt
     1621 tagttagtaa taacatcctt cataatcaac aagagttacc tacagctctt actaaattgg
     1681 ttaaagagga tgaagttgtg tcttcagaaa aagcaaaaga cagttttaat gaaaagagag
     1741 ttgcagtgga agctcctatg agggaggaat atgcagactt caaaccattt gagcgagtat
     1801 gggaagtgaa agatagtaag gaagatagtg atatgttggc tgctggaggt aaaatcgaga
     1861 gcaacttgga aagtaaagtg gataaaaaat gttttgcaga tagccttgag caaactaatc
     1921 acgaaaaaga tagtgagagt agtaatgatg atacttcttt ccccagtacg ccagaaggta
     1981 taaaggatcg ttcaggagca tatatcacat gtgctccctt taacccagca gcaactgaga
     2041 gcattgcaac aaacattttt cctttgttag gagatcctac ttcagaaaat aagaccgatg
     2101 aaaaaaaaat agaagaaaag aaggcccaaa tagtaacaga gaagaatact agcaccaaaa
     2161 catcaaaccc ttttcttgta gcagcacagg attctgagac agattatgtc acaacagata
     2221 atttaacaaa ggtgactgag gaagtcgtgg caaacatgcc tgaaggcctg actccagatt
     2281 tagtacagga agcatgtgaa agtgaattga atgaagttac tggtacaaag attgcttatg
     2341 aaacaaaaat ggacttggtt caaacatcag aagttatgca agagtcactc tatcctgcag
     2401 cacagctttg cccatcattt gaagagtcag aagctactcc ttcaccagtt ttgcctgaca
     2461 ttgttatgga agcaccattg aattctgcag ttcctagtgc tggtgcttcc gtgatacagc
     2521 ccagctcatc accattagaa gcttcttcag ttaattatga aagcataaaa catgagcctg
     2581 aaaacccccc accatatgaa gaggccatga gtgtatcact aaaaaaagta tcaggaataa
     2641 aggaagaaat taaagagcct gaaaatatta atgcagctct tcaagaaaca gaagctcctt
     2701 atatatctat tgcatgtgat ttaattaaag aaacaaagct ttctgctgaa ccagctccgg
     2761 atttctctga ttattcagaa atggcaaaag ttgaacagcc agtgcctgat cattctgagc
     2821 tagttgaaga ttcctcacct gattctgaac cagttgactt atttagtgat gattcaatac
     2881 ctgacgttcc acaaaaacaa gatgaaactg tgatgcttgt gaaagaaagt ctcactgaga
     2941 cttcatttga gtcaatgata gaatatgaaa ataaggaaaa actcagtgct ttgccacctg
     3001 agggaggaaa gccatatttg gaatctttta agctcagttt agataacaca aaagataccc
     3061 tgttacctga tgaagtttca acattgagca aaaaggagaa aattcctttg cagatggagg
     3121 agctcagtac tgcagtttat tcaaatgatg acttatttat ttctaaggaa gcacagataa
     3181 gagaaactga aacgttttca gattcatctc caattgaaat tatagatgag ttccctacat
     3241 tgatcagttc taaaactgat tcattttcta aattagccag ggaatatact gacctagaag
     3301 tatcccacaa aagtgaaatt gctaatgccc cggatggagc tgggtcattg ccttgcacag
     3361 aattgcccca tgacctttct ttgaagaaca tacaacccaa agttgaagag aaaatcagtt
     3421 tctcagatga cttttctaaa aatgggtctg ctacatcaaa ggtgctctta ttgcctccag
     3481 atgtttctgc tttggccact caagcagaga tagagagcat agttaaaccc aaagttcttg
     3541 tgaaagaagc tgagaaaaaa cttccttccg atacagaaaa agaggacaga tcaccatctg
     3601 ctatattttc agcagagctg agtaaaactt cagttgttga cctcctgtac tggagagaca
     3661 ttaagaagac tggagtggtg tttggtgcca gcctattcct gctgctttca ttgacagtat
     3721 tcagcattgt gagcgtaaca gcctacattg ccttggccct gctctctgtg accatcagct
     3781 ttaggatata caagggtgtg atccaagcta tccagaaatc agatgaaggc cacccattca
     3841 gggcatatct ggaatctgaa gttgctatat ctgaggagtt ggttcagaag tacagtaatt
     3901 ctgctcttgg tcatgtgaac tgcacgataa aggaactcag gcgcctcttc ttagttgatg
     3961 atttagttga ttctctgaag tttgcagtgt tgatgtgggt atttacctat gttggtgcct
     4021 tgtttaatgg tctgacacta ctgattttgg ctctcatttc actcttcagt gttcctgtta
     4081 tttatgaacg gcatcaggca cagatagatc attatctagg acttgcaaat aagaatgtta
     4141 aagatgctat ggctaaaatc caagcaaaaa tccctggatt gaagcgcaaa gctgaaacgg
     4201 taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gaggataaca
     4261 tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact
     4321 ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga
     4381 agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg
     4441 gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag
     4501 atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg
     4561 ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga
     4621 ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca
     4681 ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca
     4741 ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc
     4801 caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca
     4861 agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg
     4921 gcatggacga gctgtacaag taagcggccg cgactctaga tcataatcag ccataccaca
     4981 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5041 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5101 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5161 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     5221 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5281 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5341 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5401 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5461 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5521 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5581 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5641 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5701 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5761 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     5821 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5881 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5941 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6001 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6061 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6121 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     6181 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6241 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6301 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     6361 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6421 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6481 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6541 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6601 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6661 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     6721 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6781 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6841 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6901 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6961 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     7021 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     7081 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     7141 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     7201 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     7261 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     7321 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     7381 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     7441 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7501 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7561 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7621 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7681 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7741 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7801 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7861 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7921 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7981 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8041 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8101 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8161 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8221 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
