LOCUS       sequence-180319-        4999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mApple-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 4999)
  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 4999)
  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: mApple-Sec61b-C1
FEATURES             Location/Qualifiers
     source          1..4999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..73
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             364..654
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     polyA_signal    815..936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     906..925
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(943..1398)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1080..1101)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1292..1311
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1425..1529
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1531..1888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1531..1722
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1554..1574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1739..1874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1923..2717
                     /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             1923..2717
                     /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(1977..1996)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2587..2606
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2905..2924)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2949..2996
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3325..3913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3814..3833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4088..4391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4392..4595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4546..4566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4634..4643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4640..4999,1..348)
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
ORIGIN
        1 ggcgtgttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
       61 atgcagaaga agaccatggg ctgggaggcc tccgaggagc ggatgtaccc cgaggacggc
      121 gccctgaaga gcgagatcaa gaagaggctg aagctgaagg acggcggcca ctacgccgcc
      181 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc
      241 gacatcaagt tggacatcgt gtcccacaac gaggactaca ccatcgtgga acagtacgaa
      301 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga
      361 tctatgcctg gtccgacccc cagtggcact aacgtgggat cctcagggcg ctctcccagc
      421 aaagcagtgg ccgcccgggc ggcgggatcc actgtccggc agaggaaaaa tgccagctgt
      481 gggacaagga gtgcaggccg cacaacctcg gcaggcaccg gggggatgtg gcgattctac
      541 acagaagatt cacctgggct caaagttggc cctgttccag tattggttat gagtcttctg
      601 ttcatcgctt ctgtatttat gttgcacatt tggggcaagt acactcgttc gtaggaattc
      661 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
      721 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
      781 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
      841 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
      901 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
      961 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1021 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1081 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1141 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1201 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1261 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1321 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1381 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1501 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     1561 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     1621 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     1681 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     1741 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     1801 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     1861 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     1921 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     1981 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2041 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2101 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2161 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2221 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2281 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2341 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2401 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     2461 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2521 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2581 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2641 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     2701 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     2761 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     2821 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     2881 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     2941 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     3001 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3061 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3121 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     3181 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3241 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3961 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4021 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4081 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4141 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4201 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4261 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4321 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4381 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4441 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     4501 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     4561 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
     4621 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag aataacatgg ccatcatcaa
     4681 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     4741 cgagggcgag ggcgagggcc gcccctacga ggcctttcag accgctaagc tgaaggtgac
     4801 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     4861 caaggtctac attaagcacc cagccgacat ccccgactac ttcaagctgt ccttccccga
     4921 gggcttcagg tgggagcgcg tgatgaactt cgaggacggc ggcattattc acgttaacca
     4981 ggactcctcc ctgcaggac
//
