LOCUS       sequence-306136-        4964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dimerization dependent fluorescent protein RA domain fused to ER 
            protein Sec61-beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4964)
  AUTHORS   Lee JE, Cathey PI, Wu H, Parker R, Voeltz GK
  TITLE     Endoplasmic reticulum contact sites regulate the dynamics of 
            membraneless organelles.
  JOURNAL   Science. 2020 Jan 31;367(6477). pii: 367/6477/eaay7108. doi: 
            10.1126/science.aay7108.
  PUBMED    32001628
REFERENCE   2  (bases 1 to 4964)
  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: "2020-01-31"; volume: "367(6477). pii: 367/6477/eaay7108. 
            doi"; pages: " 10.1126/science.aay7108"
COMMENT     Sequence Label: RA-Sec61-beta
FEATURES             Location/Qualifiers
     source          1..4964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(366..384)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     634..653
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             947..1237
                     /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    1388..1509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1425..1444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1479..1498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1516..1971)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1653..1674)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1865..1884
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1998..2102
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2104..2461
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2104..2295
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2127..2147)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2312..2447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2496..3290
                     /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             2496..3290
                     /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(2550..2569)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3160..3179
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3478..3497)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3522..3569
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3898..4486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4387..4406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4661..4964
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatggtgag
      241 caagagcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcttgg agggctccat
      301 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
      361 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
      421 cccccagatc atgtacggct ccaaggcgta cgtgaagcac cccgccgacg tccccgatta
      481 catgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgcact tcgaggacgg
      541 cggtctggtg accgtgactc aggacacctc cctgcaggac ggcacgctga tctacaaggt
      601 gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccttggg
      661 ctgggattat gccaccgagc gcctgtaccc cgaagacggc gtgctgaagg gcgagcttct
      721 ggggcgcctg aagctgaagg acggcggcct caacctggtg gagttcaaga ccatctacat
      781 ggccaagaag cccgtgcaac tgcccggcta ctacttcgtg gacaccaagc tgggcatcac
      841 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca
      901 cctgggcatg gacgagctgt acaagtccgg actcagatct cgagctatgc ctggtccgac
      961 ccccagtggc actaacgtgg gatcctcagg gcgctctccc agcaaagcag tggccgcccg
     1021 ggcggcggga tccactgtcc ggcagaggaa aaatgccagc tgtgggacaa ggagtgcagg
     1081 ccgcacaacc tcggcaggca ccggggggat gtggcgattc tacacagaag attcacctgg
     1141 gctcaaagtt ggccctgttc cagtattggt tatgagtctt ctgttcatcg cttctgtatt
     1201 tatgttgcac atttggggca agtacactcg ttcgtagtcg tgaggtaccg cgggcccggg
     1261 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     1321 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1381 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1441 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1501 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     1561 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     1621 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     1681 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     1741 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     1801 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     1861 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     1921 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     1981 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2041 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2101 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2161 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2221 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2281 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2341 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2401 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2461 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     2521 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2581 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2641 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     2701 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2761 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     2821 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2881 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2941 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3001 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3061 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3121 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3181 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3241 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3301 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3361 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3421 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     3481 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     3541 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3601 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3661 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3721 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     3781 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3841 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4561 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     4621 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     4681 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     4741 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     4801 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     4861 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4921 ctacttggca gtacatctac gtattagtca tcgctattac catg
//
