LOCUS       sequence-179976-        4999 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses mCherry-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 Apr 14, 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: mCherry-Sec61b-C1
FEATURES             Location/Qualifiers
     source          1..4999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    403..450
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     475..494
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(682..1476)
                     /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(682..1476)
                     /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(793..812)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1403..1422
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1511..1868)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1525..1660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1677..1868
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1825..1845
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1870..1974)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2001..2456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2088..2107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2298..2319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2463..2584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2474..2493)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2528..2547
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2745..3035)
                     /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(3051..3758)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3326..3345)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3595..3613
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3755..3764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3803..4006)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3832..3852)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4007..4310
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(join(4485..4999,1..74))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4565..4584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
       61 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      121 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      181 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta
      241 tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg
      301 ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg
      361 tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac
      421 cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct
      481 ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc
      541 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat
      601 agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca
      661 tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
      721 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
      781 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
      841 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
      901 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct
      961 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     1021 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     1081 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     1141 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     1201 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     1261 cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag
     1321 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     1381 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     1441 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     1501 ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     1561 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     1621 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     1681 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     1741 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     1801 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
     1861 cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1981 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2041 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2101 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2161 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2221 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2281 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2341 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2401 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc
     2461 gttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2521 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2581 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2641 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agttatctag
     2701 atccggtgga tcccgggccc gcggtaccgt cgactgcaga attcctacga acgagtgtac
     2761 ttgccccaaa tgtgcaacat aaatacagaa gcgatgaaca gaagactcat aaccaatact
     2821 ggaacagggc caactttgag cccaggtgaa tcttctgtgt agaatcgcca catccccccg
     2881 gtgcctgccg aggttgtgcg gcctgcactc cttgtcccac agctggcatt tttcctctgc
     2941 cggacagtgg atcccgccgc ccgggcggcc actgctttgc tgggagagcg ccctgaggat
     3001 cccacgttag tgccactggg ggtcggacca ggcatagatc tgagtccgga cttgtacagc
     3061 tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg
     3121 tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc gccgggcagc
     3181 tgcacgggct tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc
     3241 ttcagcttca gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc
     3301 tcggaggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc ggaggggaag
     3361 ttggtgccgc gcagcttcac cttgtagatg aactcgccgt cctgcaggga ggagtcctgg
     3421 gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg
     3481 gggaaggaca gcttcaagta gtcggggatg tcggcggggt gcttcacgta ggccttggag
     3541 ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc acccttggtc
     3601 accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc
     3661 tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaactccttg
     3721 atgatggcca tgttatcctc ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc
     3781 ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac
     3841 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     3901 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     3961 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     4021 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     4081 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     4141 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     4201 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     4261 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     4321 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg
     4381 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4441 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4501 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4561 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4621 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4681 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4741 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4801 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4861 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4921 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4981 ggtagctctt gatccggca
//
