LOCUS       sequence-179977-        5008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mEmerald-tagged Sec61b, brightly labels endoplasmic 
            reticulum in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5008)
  AUTHORS   Nixon-Abell J, Obara CJ, Weigel AV, Li D, Legant WR, Xu CS, Pasolli 
            HA, Harvey K, Hess HF, Betzig E, Blackstone C, Lippincott-Schwartz J
  TITLE     Increased spatiotemporal resolution reveals highly dynamic dense 
            tubular matrices in the peripheral ER.
  JOURNAL   Science. 2016 Oct 28;354(6311). pii: aaf3928. Epub 2016 Oct 27.
  PUBMED    27789813
REFERENCE   2  (bases 1 to 5008)
  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"; journalName: "Science."; 
            date: "2016-10-28"; volume: "354(6311). pii"; pages: " aaf3928. Epub
            2016 Oct 27"
COMMENT     Sequence Label: mEmerald-Sec61b-C1
FEATURES             Location/Qualifiers
     source          1..5008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..198
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     438..459
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      501..510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(549..752)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(578..598)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        753..1056
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1231..1819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1311..1330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2148..2195
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2220..2239
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2427..3221)
                     /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             complement(2427..3221)
                     /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(2538..2557)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3148..3167
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3256..3613)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3270..3405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3422..3613
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3570..3590
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3615..3719)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3746..4201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3833..3852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4043..4064
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4208..4329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4219..4238)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4273..4292
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4490..4780)
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     CDS             complement(join(4796..5008,1..498,499..501,502..504))
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4838..4859)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ggtcttgaag ttcaccttga tgccgttctt ctgcttgtcg gcggtgatat agaccttgtg
       61 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
      121 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
      181 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
      241 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcac
      301 gccgtaggtc aaggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
      361 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
      421 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
      481 cagctcctcg cccttgctca ccatggtggc gaccggtagc gctagcggat ctgacggttc
      541 actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
      601 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
      661 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
      721 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
      781 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
      841 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
      901 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
      961 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     1021 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     1081 actaatgacc ccgtaattga ttactattaa taactaatgc atggcggtaa tacggttatc
     1141 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1201 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1261 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1321 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1381 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1441 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1501 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1561 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1621 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1681 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1741 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1801 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1861 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1921 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga
     1981 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     2041 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg
     2101 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     2161 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     2221 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag
     2281 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     2341 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     2401 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     2461 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     2521 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     2581 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     2641 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg
     2701 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     2761 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     2821 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     2881 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     2941 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     3001 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg
     3061 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     3121 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     3181 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     3241 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     3301 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     3361 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     3421 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     3481 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     3541 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     3601 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3661 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3721 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3781 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3841 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3901 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3961 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4021 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4081 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4141 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     4201 tacgcgttaa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4261 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4321 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     4381 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtta
     4441 tctagatccg gtggatcccg ggcccgcggt accgtcgact gcagaattcc tacgaacgag
     4501 tgtacttgcc ccaaatgtgc aacataaata cagaagcgat gaacagaaga ctcataacca
     4561 atactggaac agggccaact ttgagcccag gtgaatcttc tgtgtagaat cgccacatcc
     4621 ccccggtgcc tgccgaggtt gtgcggcctg cactccttgt cccacagctg gcatttttcc
     4681 tctgccggac agtggatccc gccgcccggg cggccactgc tttgctggga gagcgccctg
     4741 aggatcccac gttagtgcca ctgggggtcg gaccaggcat agatctgagt ccggacttgt
     4801 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     4861 gatcgcgctt ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt
     4921 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     4981 gcacgctgcc gtcctcgatg ttgtggcg
//
