LOCUS       sequence-369196-        6118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human full-length Reticulon 2 isoform B fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6118)
  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 6118)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..6118
                     /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             2005..2724
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2846..2967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2883..2902)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2937..2956
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2974..3429)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3111..3132)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3323..3342
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3456..3560
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3562..3919
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3562..3753
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3585..3605)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3770..3905
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3954..4748
                     /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             3954..4748
                     /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(4008..4027)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4618..4637
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4936..4955)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4980..5027
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5356..5944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5845..5864
                     /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 atctcgagat ggggcaggtc ctgccggtct tcgcccactg caaagaagct
      601 ccgtctacag cctcctcaac tcctgattcc acagaaggag ggaacgacga ctctgatttt
      661 cgagagctgc acacagcccg ggaattctca gaggaggacg aggaggagac cacgtcgcag
      721 gactggggca ccccccggga gctgaccttc tcctacatcg cctttgatgg tgtagtgggc
      781 tccgggggcc gcagggattc aactgcccgc cgcccccgcc cccagggccg ctcagtctcg
      841 gaaccacgag accagcaccc tcagcccagc ctgggcgaca gcttggagag catccccagc
      901 ctgagccaat ccccggagcc tggacgacgg ggtgatcctg acaccgcgcc tccatccgag
      961 cgccctctgg aagacctgag gcttcggttg gaccatctgg gctgggtggc ccggggaacg
     1021 ggatccgggg aggactcttc caccagcagc tccaccccgc tggaagacga agaaccccaa
     1081 gaacccaaca gattggagac aggagaagct ggggaagaac tggacctacg actccgactt
     1141 gctcagccct catcgcccga ggtcttgact ccccagctca gtccgggctc tgggacaccc
     1201 caggccggta ctccgtcccc atcccgatcg cgagattcga actctgggcc cgaagagcca
     1261 ttgctggaag aggaagaaaa gcagtggggg ccactggagc gagagccagt aaggggacag
     1321 tgcctcgata gcacggacca attagaattc acggtggagc cacgccttct agtggcggac
     1381 ctgctgtact ggaaggacac gaggacgtca ggagtggtct tcacaggcct gatggtctcc
     1441 ctcctctgcc tcctgcactt tagcatcgtg tccgtggccg cgcacttggc tctgttgctg
     1501 ctctgcggca ccatctctct cagggtttac cgcaaagtgc tgcaggccgt gcaccggggg
     1561 gatggagcca accctttcca ggcctacctg gatgtggacc tcaccctgac tcgggagcag
     1621 acggaacgtt tgtcccacca gatcacctcc cgcgtggtct cggcggccac gcagctgcgg
     1681 cacttcttcc tggtagaaga cctcgtggat tccctcaagc tggccctcct cttctacatc
     1741 ttgaccttcg tgggtgccat cttcaatggt ttgactcttc tcattctggg agtgattggt
     1801 ctattcacca tccccctgct gtaccggcag caccaggctc agatcgacca atatgtgggg
     1861 ttggtgacca atcagttgag ccacatcaaa gctaagatcc gagctaaaat cccagggacc
     1921 ggagccctgg cctctgcagc agccgcagtc tccggatcca aagccaaagc cgaacgggta
     1981 ccgcgggccc gggatccacc ggtcatggtg agcaagggcg ccgagctgtt caccggcatc
     2041 gtgcccatcc tgatcgagct gaatggcgat gtgaatggcc acaagttcag cgtgagcggc
     2101 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2161 aagctgcctg tgccctggcc caccctggtg accaccctga gctacggcgt gcagtgcttc
     2221 tcacgctacc ccgatcacat gaagcagcac gacttcttca agagcgccat gcctgagggc
     2281 tacatccagg agcgcaccat cttcttcgag gatgacggca actacaagtc gcgcgccgag
     2341 gtgaagttcg agggcgatac cctggtgaat cgcatcgagc tgaccggcac cgatttcaag
     2401 gaggatggca acatcctggg caataagatg gagtacaact acaacgccca caatgtgtac
     2461 atcatgaccg acaaggccaa gaatggcatc aaggtgaact tcaagatccg ccacaacatc
     2521 gaggatggca gcgtgcagct ggccgaccac taccagcaga atacccccat cggcgatggc
     2581 cctgtgctgc tgcccgataa ccactacctg tccacccaga gcgccctgtc caaggacccc
     2641 aacgagaagc gcgatcacat gatctacttc ggcttcgtga ccgccgccgc catcacccac
     2701 ggcatggatg agctgtacaa gtgagcggcc gcgactctag atcataatca gccataccac
     2761 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2821 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2881 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2941 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     3001 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3061 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3121 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3181 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3241 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3301 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3361 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3421 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3481 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3541 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3601 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3661 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3721 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3781 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3841 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3901 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3961 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4021 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4081 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4141 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4201 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4261 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4321 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4381 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4441 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4501 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4561 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4621 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4681 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4741 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4801 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4861 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4921 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4981 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5041 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5101 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5161 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5221 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5281 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5401 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5461 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5521 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5581 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5641 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5701 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5761 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5821 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5881 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5941 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6001 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     6061 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
