LOCUS       sequence-227923-        4984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Sec61 beta fused to TagBFP, used as a general ER 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4984)
  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 4984)
  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: BFP-Sec61 beta
FEATURES             Location/Qualifiers
     source          1..4984
                     /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             614..1312
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1328..1618
                     /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    1779..1900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1816..1835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1870..1889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1907..2362)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2044..2065)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2256..2275
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2389..2493
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2495..2852
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2495..2686
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2518..2538)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2703..2838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2887..3681
                     /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             2887..3681
                     /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(2941..2960)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3551..3570
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3869..3888)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3913..3960
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4289..4877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4778..4797
                     /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 agcggtcgcc accatgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
      661 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
      721 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
      781 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
      841 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
      901 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
      961 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     1021 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     1081 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac
     1141 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     1201 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     1261 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta attccggact
     1321 cagatctatg cctggtccga cccccagtgg cactaacgtg ggatcctcag ggcgctctcc
     1381 cagcaaagca gtggccgccc gggcggcggg atccactgtc cggcagagga aaaatgccag
     1441 ctgtgggaca aggagtgcag gccgcacaac ctcggcaggc accgggggga tgtggcgatt
     1501 ctacacagaa gattcacctg ggctcaaagt tggccctgtt ccagtattgg ttatgagtct
     1561 tctgttcatc gcttctgtat ttatgttgca catttggggc aagtacactc gttcgtagga
     1621 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     1681 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1741 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1801 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1861 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     1921 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1981 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2041 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2101 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2161 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2221 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2281 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2341 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2401 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2461 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2521 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2581 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2641 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2701 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2761 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2821 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2881 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2941 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3001 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3061 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3121 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3181 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3241 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3301 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3361 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3421 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3481 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3541 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3601 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3661 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3721 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3781 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3841 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3901 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     3961 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4021 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4081 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4141 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4201 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4261 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4681 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4921 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4981 ccgc
//
