LOCUS       sequence-338349-        4986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  General ER marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4986)
  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 4986)
  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: mNeonGreen-Sec61-beta
FEATURES             Location/Qualifiers
     source          1..4986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             604..1311
                     /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      859..868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1342..1629
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="PGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAGR
                     TTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     polyA_signal    1774..1895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1811..1830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1865..1884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1902..2357)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2039..2060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2251..2270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2384..2488
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2490..2847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2490..2681
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2513..2533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2698..2833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2882..3676
                     /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             2882..3676
                     /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(2936..2955)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3546..3565
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3864..3883)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3908..3955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4284..4872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4773..4792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc
      601 accatggtga gcaagggcga ggaggataac atggcctctc tcccagcgac acatgagtta
      661 cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg caccggcaat
      721 ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct ccagttctcc
      781 ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc ctaccctgac
      841 gggatgtcgc ctttccaggc cgccatggta gatggctccg gctaccaagt ccatcgcaca
      901 atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta cgagggaagc
      961 cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg tcctgtgatg
     1021 accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc caacgacaaa
     1081 accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg ctaccggagc
     1141 actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct gaagaaccag
     1201 ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct caacttcaag
     1261 gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa gtccggactc
     1321 agatctcgag ctcaagcttc gcctggtccg acccccagtg gcactaacgt gggatcctca
     1381 gggcgctctc ccagcaaagc agtggccgcc cgggcggcgg gatccactgt ccggcagagg
     1441 aaaaatgcca gctgtgggac aaggagtgca ggccgcacaa cctcggcagg caccgggggg
     1501 atgtggcgat tctacacaga agattcacct gggctcaaag ttggccctgt tccagtattg
     1561 gttatgagtc ttctgttcat cgcttctgta tttatgttgc acatttgggg caagtacact
     1621 cgttcgtagg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     1681 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1741 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1801 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1861 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     1921 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1981 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2041 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2101 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2161 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2221 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2281 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2341 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2401 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2461 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2521 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2581 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2641 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2701 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2761 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2821 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     2881 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2941 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3001 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3061 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3121 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3181 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3241 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3301 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3361 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3421 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3481 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3541 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3601 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3661 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3721 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3781 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3841 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3901 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     3961 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4021 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4081 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4141 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4201 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4261 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4321 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4381 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4441 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4501 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4561 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4621 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4681 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4741 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4801 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4861 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4921 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4981 atgcat
//
