LOCUS       sequence-369195-        5512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human Reticulon 2 isoform B N-terminal domain aa 1 to 271 
            fused to mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5512)
  AUTHORS   Zamponi E, Meehl JB, Voeltz GK
  TITLE     The ER ladder is a unique morphological feature of developing 
            mammalian axons.
  JOURNAL   Dev Cell. 2022 Jun 6;57(11):1369-1382.e6. doi: 
            10.1016/j.devcel.2022.05.002. Epub 2022 May 23.
  PUBMED    35609616
REFERENCE   2  (bases 1 to 5512)
  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.2022.05.002"; journalName: "Dev Cell"; date: 
            "2022-06-6- 6"; volume: "57"; issue: "11"; pages: "1369-1382.e6"
COMMENT     Sequence Label: Rtn2b-NTD1-271-mNeon
FEATURES             Location/Qualifiers
     source          1..5512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      845..854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             851..1561
                     /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      1106..1115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1683..1804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1720..1739)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1774..1793
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1811..2266)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1948..1969)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2160..2179
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2293..2397
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2399..2756
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2399..2590
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2422..2442)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2607..2742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2791..3585
                     /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             2791..3585
                     /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(2845..2864)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3455..3474
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3773..3792)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3817..3864
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4193..4781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4682..4701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4956..5259
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5260..5463
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5414..5434
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gatggggcag gtcctgccgg tcttcgccca ctgcaaagaa gctccgtcta cagcctcctc
       61 aactcctgat tccacagaag gagggaacga cgactctgat tttcgagagc tgcacacagc
      121 ccgggaattc tcagaggagg acgaggagga gaccacgtcg caggactggg gcaccccccg
      181 ggagctgacc ttctcctaca tcgcctttga tggtgtagtg ggctccgggg gccgcaggga
      241 ttcaactgcc cgccgccccc gcccccaggg ccgctcagtc tcggaaccac gagaccagca
      301 ccctcagccc agcctgggcg acagcttgga gagcatcccc agcctgagcc aatccccgga
      361 gcctggacga cggggtgatc ctgacaccgc gcctccatcc gagcgccctc tggaagacct
      421 gaggcttcgg ttggaccatc tgggctgggt ggcccgggga acgggatccg gggaggactc
      481 ttccaccagc agctccaccc cgctggaaga cgaagaaccc caagaaccca acagattgga
      541 gacaggagaa gctggggaag aactggacct acgactccga cttgctcagc cctcatcgcc
      601 cgaggtcttg actccccagc tcagtccggg ctctgggaca ccccaggccg gtactccgtc
      661 cccatcccga tcgcgagatt cgaactctgg gcccgaagag ccattgctgg aagaggaaga
      721 aaagcagtgg gggccactgg agcgagagcc agtaagggga cagtgcctcg atagcacgga
      781 ccaattagaa ttcacggtgg agccacgcct tctacgggta ccgcgggccc gggatccacc
      841 ggtcgccacc atggtgagca agggcgagga ggataacatg gcctctctcc cagcgacaca
      901 tgagttacac atctttggct ccatcaacgg tgtggacttt gacatggtgg gtcagggcac
      961 cggcaatcca aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca
     1021 gttctccccc tggattctgg tccctcatat cgggtatggc ttccatcagt acctgcccta
     1081 ccctgacggg atgtcgcctt tccaggccgc catggtagat ggctccggct accaagtcca
     1141 tcgcacaatg cagtttgaag atggtgcctc ccttactgtt aactaccgct acacctacga
     1201 gggaagccac atcaaaggag aggcccaggt gaaggggact ggtttccctg ctgacggtcc
     1261 tgtgatgacc aactcgctga ccgctgcgga ctggtgcagg tcgaagaaga cttaccccaa
     1321 cgacaaaacc atcatcagta cctttaagtg gagttacacc actggaaatg gcaagcgcta
     1381 ccggagcact gcgcggacca cctacacctt tgccaagcca atggcggcta actatctgaa
     1441 gaaccagccg atgtacgtgt tccgtaagac ggagctcaag cactccaaga ccgagctcaa
     1501 cttcaaggag tggcaaaagg cctttaccga tgtgatgggc atggacgagc tgtacaagta
     1561 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1621 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1681 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1741 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1801 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     1861 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     1921 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     1981 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2041 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2101 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2161 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2221 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     2281 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2341 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     2401 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2581 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2641 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2701 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     2761 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2821 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2881 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2941 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3001 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3061 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3121 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3181 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3241 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3301 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3361 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3421 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3481 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3541 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3601 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3661 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3721 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     3781 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3841 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     3901 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     3961 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4021 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4081 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4141 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4201 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4261 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4321 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4381 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4441 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4501 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4561 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4621 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4681 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4741 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4801 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4861 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     4921 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     4981 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5041 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5101 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5161 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5221 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     5281 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     5341 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     5401 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     5461 gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc ga
//
