LOCUS       sequence-338353-        7723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Rtn3L with 6x LC3-interacting regions mutated in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7723)
  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 7723)
  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: Rtn3L-Delta-6LIR-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..7723
                     /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             complement(1023..1034)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3673..3682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3679..4389
                     /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      3934..3943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4511..4632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4548..4567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4602..4621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4639..5094)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4776..4797)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4988..5007
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5121..5225
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5227..5584
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5227..5418
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5250..5270)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5435..5570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5619..6413
                     /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             5619..6413
                     /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(5673..5692)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6283..6302
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6601..6620)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6645..6692
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7021..7609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7510..7529
                     /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 agggattgag ctctctttgc
      781 tctgatgagc catcttcaga aattatgact tcttcctttc tttcatcttc tgaaatacat
      841 aacactggcc ttacaatact acatggagaa aaaagccatg tgttagggag ccagcctatt
      901 ttagccaaag aaggaaaaga ccacttggat cttctagata tgaaaaagat ggaaaagcct
      961 caggggacca gcaacaacgt atcagactct tcagtttctc ttgcagcagg agttcattgt
     1021 gaccgtcctt ctattccagc cagtttccca gagcatcctg cttttctctc aaagaaaatt
     1081 ggtcaagtgg aagagcaaat agataaagag accaagaacc caaatggggt atcaagtagg
     1141 gaggctaaaa ctgcattgga tgctgatgac agagccactt tggcgacagc ccagaaacca
     1201 cctactgagg cctctaaggc agaaggcatt tatacatatt ctttgtctcc atccaaagtt
     1261 tcaggagatg atgttattga aaaggattcc cctgaatcac cagctgaagt agctattgac
     1321 aaagcagcat ttgacaaaga atttaaagac tcatataagg agagcacaga tgattttggt
     1381 agctggtctg tgcacactga taaagaatca tccgaagaca tttcagagac taatgacaag
     1441 ctttttccac tgagaaataa agaggcagga cgttacccaa tgtctgcatt gctcagtagg
     1501 cagttttcac acacaaatgc agcactggaa gaggtgtcca gatgcgtgaa tgatatgcat
     1561 aactttacta acgaaatact gactgcggat ctggctcccc aagtgaaaca acagaccgat
     1621 aaatcttctg actgcatcac aaaaactaca ggacttgaca tgagtgaata taattcagaa
     1681 attccagttg taaatcttaa aactagcact catcagaaaa ctcctgtatg ttctattgat
     1741 gggagcactc ccatcactaa atcaacaggt gattgggcag aagcatctct ccagcaagaa
     1801 aatgctatta ctggaaaacc tgtacctgac tctttgaatt ccacaaaaga attcagtatc
     1861 aaaggtgtgc aaggcaatat gcagaaacag gatgacacac ttgcagaatt acctggatct
     1921 ccacctgaga aatgtgactc tttgggttct ggagtggcca cagtgaaagt ggttttacct
     1981 gatgaccacc tgaaagatga aatggactgg cagagctctg cattgggaga aatcacagaa
     2041 gctgatagtt ctggtgagtc tgatgacaca gtaatagagg acatcacagc agatacatca
     2101 tttgaaaata acaaaattca ggctgaaaaa cctgtttcca ttccaagtgc tgttgtaaaa
     2161 acaggtgaaa gagaaatcaa agagattccc agttgtgaga gagaagaaaa aacatctaaa
     2221 aacgctgaag aagcggtcag tgactctgag ctgcatcaag atcagcctga tattcttgga
     2281 aggagtccag ctagtgaggc agcatgttca aaagtacccg atacgaatgt ctccttagaa
     2341 gatgtgagtg aagttgctcc tgaaaagcct attactactg agaaccccaa acttccttca
     2401 acagtgtctc caaatgtttt taatgagaca gaattctcat taaatgtgac aacatctgcc
     2461 tatttggagt cattacatgg gaaaaatgtt aaacatatag atgattcctc cccagaggac
     2521 ctgatagcag cctttacaga aaccagagat aaaggaatag tagatagtga aagaaatgct
     2581 tttaaagcaa tatcagagaa gatgacagac tttaaaacaa ctcctcctgt agaagtctta
     2641 catgaaaatg agtccggtgg ttctgaaatt aaagacattg gaagcaaata cagtgaacaa
     2701 agcaaagaaa caaatggaag tgagcctcta ggtgttttcc ctacccaagg tactccagta
     2761 gcatctcttg acttagaaca agaacagctc acaattaagg ctcttaaaga attaggtgaa
     2821 agacaggttg agaagtcaac ttctgcacag cgtgacgcag aattgccttc tgaagaagta
     2881 ctgaagcaaa ctttcacatt tgctccagaa tcttggccac agagatcagc tgacatcgca
     2941 gaacgtaatg tcaagaatgg atctgatctt gggatttccc agaagcccat cactatcaga
     3001 gaaactacta gggtagatgc tgtttccagc cttagcaaga ctgaattggt aaaaaagcat
     3061 gtcctagcaa gacttctgac agacttctca gtgcacgatc tgattttctg gagagatgtg
     3121 aagaagactg ggtttgtctt tggcaccacg ctgatcatgc tgctttccct ggcagctttc
     3181 agtgtcatca gtgtggtttc ttacctcatc ctggctcttc tctctgtcac catcagcttc
     3241 aggatctaca agtccgtcat ccaagctgta cagaagtcag aagaaggcca tccattcaaa
     3301 gcctacctgg acgtagacat tactctgtcc tcagaagctt tccataatta catgaatgct
     3361 gccatggtgc acatcaacag ggccctgaaa ctcattattc gtctctttct ggtagaagat
     3421 ctggttgact ccttgaagct ggctgtcttc atgtggctga tgacctatgt tggtgctgtt
     3481 tttaacggaa tcacccttct aattcttgct gaactgctca ttttcagtgt cccgattgtc
     3541 tatgagaagt acaagaccca gattgatcac tatgttggca tcgcccgaga tcagaccaag
     3601 tcaattgttg aaaagatcca agcaaaactc cctggaatcg ccaaaaaaaa ggcagaacgg
     3661 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc ctctctccca
     3721 gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt
     3781 cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt
     3841 gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac
     3901 ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggctac
     3961 caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac
     4021 acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct
     4081 gacggtcctg tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact
     4141 taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc
     4201 aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac
     4261 tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc
     4321 gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg
     4381 tacaagtaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     4441 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4501 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4561 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4621 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4681 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4741 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4801 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc tatggcccac
     4861 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4921 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4981 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5041 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     5101 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5161 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5221 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5341 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5401 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5461 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5521 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5581 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5641 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5701 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5761 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5821 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5881 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5941 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6001 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6061 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6121 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     6181 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6241 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6301 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6361 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6421 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6481 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6541 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     6601 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6661 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6721 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6781 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6841 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6901 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6961 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7021 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7081 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7141 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7201 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7261 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7321 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7381 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7441 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7501 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7561 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7621 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7681 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
