LOCUS       sequence-369198-        5112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human Reticulon 2 isoform B N-terminal domain aa 136 to 271 
            fused to mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5112)
  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 5112)
  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-NTD136-271-mNeon
FEATURES             Location/Qualifiers
     source          1..5112
                     /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"
     regulatory      1002..1011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1008..1718
                     /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      1263..1272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1840..1961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1877..1896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1931..1950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1968..2423)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2105..2126)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2317..2336
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2450..2554
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2556..2913
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2556..2747
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2579..2599)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2764..2899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2948..3742
                     /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             2948..3742
                     /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(3002..3021)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3612..3631
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3930..3949)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3974..4021
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4350..4938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4839..4858
                     /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 atgcctctgg aagacctgag gcttcggttg gaccatctgg
      601 gctgggtggc ccggggaacg ggatccgggg aggactcttc caccagcagc tccaccccgc
      661 tggaagacga agaaccccaa gaacccaaca gattggagac aggagaagct ggggaagaac
      721 tggacctacg actccgactt gctcagccct catcgcccga ggtcttgact ccccagctca
      781 gtccgggctc tgggacaccc caggccggta ctccgtcccc atcccgatcg cgagattcga
      841 actctgggcc cgaagagcca ttgctggaag aggaagaaaa gcagtggggg ccactggagc
      901 gagagccagt aaggggacag tgcctcgata gcacggacca attagaattc acggtggagc
      961 cacgccttct acgggtaccg cgggcccggg atccaccggt cgccaccatg gtgagcaagg
     1021 gcgaggagga taacatggcc tctctcccag cgacacatga gttacacatc tttggctcca
     1081 tcaacggtgt ggactttgac atggtgggtc agggcaccgg caatccaaat gatggttatg
     1141 aggagttaaa cctgaagtcc accaagggtg acctccagtt ctccccctgg attctggtcc
     1201 ctcatatcgg gtatggcttc catcagtacc tgccctaccc tgacgggatg tcgcctttcc
     1261 aggccgccat ggtagatggc tccggctacc aagtccatcg cacaatgcag tttgaagatg
     1321 gtgcctccct tactgttaac taccgctaca cctacgaggg aagccacatc aaaggagagg
     1381 cccaggtgaa ggggactggt ttccctgctg acggtcctgt gatgaccaac tcgctgaccg
     1441 ctgcggactg gtgcaggtcg aagaagactt accccaacga caaaaccatc atcagtacct
     1501 ttaagtggag ttacaccact ggaaatggca agcgctaccg gagcactgcg cggaccacct
     1561 acacctttgc caagccaatg gcggctaact atctgaagaa ccagccgatg tacgtgttcc
     1621 gtaagacgga gctcaagcac tccaagaccg agctcaactt caaggagtgg caaaaggcct
     1681 ttaccgatgt gatgggcatg gacgagctgt acaagtaagc ggccgcgact ctagatcata
     1741 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1801 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     1861 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1921 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     1981 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2041 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2101 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2161 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2221 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2281 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2341 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2401 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2461 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2521 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     2581 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2641 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2701 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2761 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2821 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2881 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     2941 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3001 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3061 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3121 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3181 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3241 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3301 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3361 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3421 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     3481 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3541 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3601 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3661 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3721 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3781 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3841 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3901 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     3961 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4021 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4081 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4141 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4201 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4261 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4321 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4381 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4441 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4501 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4561 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4621 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4681 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4741 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4801 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4861 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4921 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4981 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5041 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5101 tatggagttc cg
//
