LOCUS       sequence-338188-        5395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Rtn3S in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5395)
  AUTHORS   Wu H, Voeltz GK
  TITLE     Reticulon-3 Promotes Endosome Maturation at ER Membrane Contact 
            Sites.
  JOURNAL   Dev Cell. 2021 Jan 11;56(1):52-66.e7. doi: 
            10.1016/j.devcel.2020.12.014.
  PUBMED    33434526
REFERENCE   2  (bases 1 to 5395)
  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.2020.12"; journalName: "Dev Cell"; date: 
            "2021-01-11- 11"; volume: "56"; issue: "1"; pages: "52-66.e7"
COMMENT     Sequence Label: Rtn3S-GFP
FEATURES             Location/Qualifiers
     source          1..5395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1342..2061
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2183..2304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2220..2239)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2274..2293
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2311..2766)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2448..2469)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2660..2679
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2793..2897
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2899..3256
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2899..3090
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2922..2942)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3107..3242
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3291..4085
                     /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             3291..4085
                     /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(3345..3364)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3955..3974
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4273..4292)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4317..4364
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4693..5281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5182..5201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagat ggcggagccg tcggcggcca ctcagtccca ttccatctcc
      661 tcgtcgtcct tcggagccga gccgtccgcg cccggcggcg gcgggagccc aggagcctgc
      721 cccgccctgg ggacgaagag ctgcagctcc tcctgtgcgg tgcacgatct gattttctgg
      781 agagatgtga agaagactgg gtttgtcttt ggcaccacgc tgatcatgct gctttccctg
      841 gcagctttca gtgtcatcag tgtggtttct tacctcatcc tggctcttct ctctgtcacc
      901 atcagcttca ggatctacaa gtccgtcatc caagctgtac agaagtcaga agaaggccat
      961 ccattcaaag cctacctgga cgtagacatt actctgtcct cagaagcttt ccataattac
     1021 atgaatgctg ccatggtgca catcaacagg gccctgaaac tcattattcg tctctttctg
     1081 gtagaagatc tggttgactc cttgaagctg gctgtcttca tgtggctgat gacctatgtt
     1141 ggtgctgttt ttaacggaat cacccttcta attcttgctg aactgctcat tttcagtgtc
     1201 ccgattgtct atgagaagta caagacccag attgatcact atgttggcat cgcccgagat
     1261 cagaccaagt caattgttga aaagatccaa gcaaaactcc ctggaatcgc caaaaaaaag
     1321 gcagaacggg atccaccggt catggtgagc aagggcgccg agctgttcac cggcatcgtg
     1381 cccatcctga tcgagctgaa tggcgatgtg aatggccaca agttcagcgt gagcggcgag
     1441 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1501 ctgcctgtgc cctggcccac cctggtgacc accctgagct acggcgtgca gtgcttctca
     1561 cgctaccccg atcacatgaa gcagcacgac ttcttcaaga gcgccatgcc tgagggctac
     1621 atccaggagc gcaccatctt cttcgaggat gacggcaact acaagtcgcg cgccgaggtg
     1681 aagttcgagg gcgataccct ggtgaatcgc atcgagctga ccggcaccga tttcaaggag
     1741 gatggcaaca tcctgggcaa taagatggag tacaactaca acgcccacaa tgtgtacatc
     1801 atgaccgaca aggccaagaa tggcatcaag gtgaacttca agatccgcca caacatcgag
     1861 gatggcagcg tgcagctggc cgaccactac cagcagaata cccccatcgg cgatggccct
     1921 gtgctgctgc ccgataacca ctacctgtcc acccagagcg ccctgtccaa ggaccccaac
     1981 gagaagcgcg atcacatgat ctacttcggc ttcgtgaccg ccgccgccat cacccacggc
     2041 atggatgagc tgtacaagtg agcggccgcg actctagatc ataatcagcc ataccacatt
     2101 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2161 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2221 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2281 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2341 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2401 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2461 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2521 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2581 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2641 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2701 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2761 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2881 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2941 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3001 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3061 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3121 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3181 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3241 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3301 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3361 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3421 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3481 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3541 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3601 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3661 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3721 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3781 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     3841 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3901 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3961 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4021 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4081 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4141 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4201 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4261 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4321 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4381 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4441 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4501 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4561 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4621 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4681 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4741 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4801 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4861 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4921 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4981 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5041 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5101 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5161 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5221 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5281 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5341 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
