LOCUS       sequence-116488-        5699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Sec61 beta fused to mCherry, used as a general ER 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5699)
  AUTHORS   Zurek N, Sparks L, Voeltz G
  TITLE     Reticulon short hairpin transmembrane domains are used to shape ER 
            tubules.
  JOURNAL   Traffic. 2011 Jan;12(1):28-41. doi: 
            10.1111/j.1600-0854.2010.01134.x. Epub 2010 Nov 12.
  PUBMED    20955502
REFERENCE   2  (bases 1 to 5699)
  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: 
            "10.1111/j.1600-0854.2010.01134.x"; journalName: "Traffic"; date: 
            "2011-01"; volume: "12"; issue: "1"; pages: "28-41"
COMMENT     Sequence Label: mCh-Sec61 beta
FEATURES             Location/Qualifiers
     source          1..5699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             604..1311
                     /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(749..767)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1017..1036
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1318..1608
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     CDS             1653..2372
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2494..2615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2531..2550)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2585..2604
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2622..3077)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2759..2780)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2971..2990
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3104..3208
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3210..3567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3210..3401
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3233..3253)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3418..3553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3602..4396
                     /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             3602..4396
                     /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(3656..3675)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4266..4285
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4584..4603)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4628..4675
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5004..5592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5493..5512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
      661 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      721 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
      781 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
      841 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
      901 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      961 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1021 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1081 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     1141 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     1201 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1261 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gagatctatg
     1321 cctggtccga cccccagtgg cactaacgtg ggatcctcag ggcgctctcc cagcaaagca
     1381 gtggccgccc gggcggcggg atccactgtc cggcagagga aaaatgccag ctgtgggaca
     1441 aggagtgcag gccgcacaac ctcggcaggc accgggggga tgtggcgatt ctacacagaa
     1501 gattcacctg ggctcaaagt tggccctgtt ccagtattgg ttatgagtct tctgttcatc
     1561 gcttctgtat ttatgttgca catttggggc aagtacactc gttcgtagga attctgcagt
     1621 cgacggtacc gcgggcccgg gatccaccgg tcatggtgag caagggcgcc gagctgttca
     1681 ccggcatcgt gcccatcctg atcgagctga atggcgatgt gaatggccac aagttcagcg
     1741 tgagcggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1801 ccaccggcaa gctgcctgtg ccctggccca ccctggtgac caccctgagc tacggcgtgc
     1861 agtgcttctc acgctacccc gatcacatga agcagcacga cttcttcaag agcgccatgc
     1921 ctgagggcta catccaggag cgcaccatct tcttcgagga tgacggcaac tacaagtcgc
     1981 gcgccgaggt gaagttcgag ggcgataccc tggtgaatcg catcgagctg accggcaccg
     2041 atttcaagga ggatggcaac atcctgggca ataagatgga gtacaactac aacgcccaca
     2101 atgtgtacat catgaccgac aaggccaaga atggcatcaa ggtgaacttc aagatccgcc
     2161 acaacatcga ggatggcagc gtgcagctgg ccgaccacta ccagcagaat acccccatcg
     2221 gcgatggccc tgtgctgctg cccgataacc actacctgtc cacccagagc gccctgtcca
     2281 aggaccccaa cgagaagcgc gatcacatga tctacttcgg cttcgtgacc gccgccgcca
     2341 tcacccacgg catggatgag ctgtacaagt gagcggccgc gactctagat cataatcagc
     2401 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2461 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2521 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2581 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     2641 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2701 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2761 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2821 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2881 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2941 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3001 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3061 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3181 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3241 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3301 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3361 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3421 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3481 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3541 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3601 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3661 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3721 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3781 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3841 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3901 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3961 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4021 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4081 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4141 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4201 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4261 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4321 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4381 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4441 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4501 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4561 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4621 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4681 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4741 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4801 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4861 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4921 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4981 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5041 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5101 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5161 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5221 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5281 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5341 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5401 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5461 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5521 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5581 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5641 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgc
//
