LOCUS       sequence-369206-        4870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human Reticulon 2 disease mutant R60f fused to BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4870)
  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 4870)
  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-R60f-BFP
FEATURES             Location/Qualifiers
     source          1..4870
                     /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             775..1476
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    1598..1719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1635..1654)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1689..1708
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1726..2181)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1863..1884)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2075..2094
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2208..2312
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2314..2671
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2314..2505
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2337..2357)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2522..2657
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2706..3500
                     /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             2706..3500
                     /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(2760..2779)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3370..3389
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3688..3707)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3732..3779
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4108..4696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4597..4616
                     /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 cccccccggg agctgacctt ctcctacatc gccttgcacc ggtcatgagc
      781 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat
      841 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
      901 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc
      961 ctctacggca gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag
     1021 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     1081 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     1141 ggggtgaact tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     1201 ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg
     1261 aagctcgtgg gcgggagcca tctgatcgca aacatcaaga ccacatatag atccaagaaa
     1321 cccgctaaga acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc
     1381 aaggaggcca acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc
     1441 gacctcccta gcaaactggg gcacaagctt aattgagcgg ccgcgactct agatcataat
     1501 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1561 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1621 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1681 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     1741 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1801 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1861 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1921 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1981 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2041 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2101 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2161 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2221 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2281 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2341 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2401 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2461 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2521 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2581 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2641 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     2701 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2761 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2821 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2881 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     2941 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3001 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3061 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3121 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3181 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3241 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3301 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3361 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3421 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3481 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3541 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3601 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3661 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3721 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     3781 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3841 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3901 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     3961 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4021 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4081 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4141 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4201 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4261 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4321 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4381 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4441 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4501 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4561 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4621 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4681 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4741 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4801 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4861 tggagttccg
//
