LOCUS       sequence-369199-        5317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human Reticulon 2 isoform B reticulon homology domain aa 272
            to 472 fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5317)
  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 5317)
  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-RHD272-472-GFP
FEATURES             Location/Qualifiers
     source          1..5317
                     /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             1204..1923
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2045..2166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2082..2101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2136..2155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2173..2628)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2310..2331)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2522..2541
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2655..2759
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2761..3118
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2761..2952
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2784..2804)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2969..3104
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3153..3947
                     /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             3153..3947
                     /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(3207..3226)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3817..3836
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4135..4154)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4179..4226
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4555..5143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5044..5063
                     /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 ggggagtaaa gtggcggacc tgctgtactg gaaggacacg
      601 aggacgtcag gagtggtctt cacaggcctg atggtctccc tcctctgcct cctgcacttt
      661 agcatcgtgt ccgtggccgc gcacttggct ctgttgctgc tctgcggcac catctctctc
      721 agggtttacc gcaaagtgct gcaggccgtg caccgggggg atggagccaa ccctttccag
      781 gcctacctgg atgtggacct caccctgact cgggagcaga cggaacgttt gtcccaccag
      841 atcacctccc gcgtggtctc ggcggccacg cagctgcggc acttcttcct ggtagaagac
      901 ctcgtggatt ccctcaagct ggccctcctc ttctacatct tgaccttcgt gggtgccatc
      961 ttcaatggtt tgactcttct cattctggga gtgattggtc tattcaccat ccccctgctg
     1021 taccggcagc accaggctca gatcgaccaa tatgtggggt tggtgaccaa tcagttgagc
     1081 cacatcaaag ctaagatccg agctaaaatc ccagggaccg gagccctggc ctctgcagca
     1141 gccgcagtct ccggatccaa agccaaagcc gaacgggtac cgcgggcccg ggatccaccg
     1201 gtcatggtga gcaagggcgc cgagctgttc accggcatcg tgcccatcct gatcgagctg
     1261 aatggcgatg tgaatggcca caagttcagc gtgagcggcg agggcgaggg cgatgccacc
     1321 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcctgt gccctggccc
     1381 accctggtga ccaccctgag ctacggcgtg cagtgcttct cacgctaccc cgatcacatg
     1441 aagcagcacg acttcttcaa gagcgccatg cctgagggct acatccagga gcgcaccatc
     1501 ttcttcgagg atgacggcaa ctacaagtcg cgcgccgagg tgaagttcga gggcgatacc
     1561 ctggtgaatc gcatcgagct gaccggcacc gatttcaagg aggatggcaa catcctgggc
     1621 aataagatgg agtacaacta caacgcccac aatgtgtaca tcatgaccga caaggccaag
     1681 aatggcatca aggtgaactt caagatccgc cacaacatcg aggatggcag cgtgcagctg
     1741 gccgaccact accagcagaa tacccccatc ggcgatggcc ctgtgctgct gcccgataac
     1801 cactacctgt ccacccagag cgccctgtcc aaggacccca acgagaagcg cgatcacatg
     1861 atctacttcg gcttcgtgac cgccgccgcc atcacccacg gcatggatga gctgtacaag
     1921 tgagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1981 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2041 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2101 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2161 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2221 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2281 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2341 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2401 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2461 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2521 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2581 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     2641 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2701 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     2761 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2821 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2881 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2941 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3001 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3061 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3121 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3181 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3241 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3301 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     3361 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3421 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3481 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3541 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3601 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3661 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3721 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3781 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3841 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3901 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3961 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4021 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4081 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4141 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4201 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4261 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4321 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4381 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     4441 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4501 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4561 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4621 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4681 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4741 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4801 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4861 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4921 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4981 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5041 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5101 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5161 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5221 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     5281 cggggtcatt agttcatagc ccatatatgg agttccg
//
