LOCUS       sequence-352455-        4718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  visualization of the ER.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4718)
  AUTHORS   Friedman JR, Lackner LL, West M, DiBenedetto JR, Nunnari J, Voeltz 
            GK
  TITLE     ER tubules mark sites of mitochondrial division.
  JOURNAL   Science. 2011 Oct 21;334(6054):358-62. doi: 10.1126/science.1207385.
            Epub 2011 Sep 1.
  PUBMED    21885730
REFERENCE   2  (bases 1 to 4718)
  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: 
            "10.1126/science.1207385"; journalName: "Science"; date: 
            "2011-10-21- 21"; volume: "334"; issue: "6054"; pages: "358-62"
COMMENT     Sequence Label: BFP-KDEL
FEATURES             Location/Qualifiers
     source          1..4718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(10..367)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      24..159
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        176..367
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     324..344
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(369..473)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      500..955
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(587..606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     797..818
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    962..1083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(973..992)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1027..1046
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1243..1938)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     promoter        complement(2021..2224)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2050..2070)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2225..2528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2703..3291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2783..2802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3620..3667
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3692..3711
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3899..4693)
                     /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(3899..4693)
                     /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(4010..4029)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4620..4639
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
       61 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
      121 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
      181 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
      241 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      301 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
      361 gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      421 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      481 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      541 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      721 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      781 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      841 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      901 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacgcg
      961 ttaagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1021 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1081 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1141 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca gttatctaga
     1201 tccggtggat cccgggcccg cggtacctca tagctcgtct ttattaagct tgtgccccag
     1261 tttgctaggg aggtcgcagt atctggccac tgccacctcg tgctgctcga cgtaggtctc
     1321 gttgttggcc tccttgattc tttccagtct gtagtccaca tagtagacgc caggcatctt
     1381 gaggttctta gcgggtttct tggatctata tgtggtcttg atgtttgcga tcagatggct
     1441 cccgcccacg agcttcaggg ccatgtcgtt tctgccttcc aggccgccgt cagcggggta
     1501 cagcgtctcg gtgaaggcct cccagccgag tgttttcttc tgcatcacag ggccgttgga
     1561 tgtgaagttc acccctctga tcttgacgtt gtagatgagg cagccgtcct ggaggctggt
     1621 gtcctgggta gcggtcagca cgcccccgtc ttcgtatgtg gtgactctct cccatgtgaa
     1681 gccctcaggg aaggactgct tgaagaagtc ggggatgccc tgggtgtggt tgatgaaggt
     1741 cttgctgccg tagaggaagc tagtagccag gatgtcgaag gcgaagggga gagggccgcc
     1801 ctcgaccacc ttgattctca tggtctgggt gccctcgtag ggcttgcctt cgccctcgga
     1861 tgtgcacttg aagtgatggt tgtccacggt gccctccatg tacagcttca tgtgcatgtt
     1921 ctccttaatc agctcgctgg cccgcgccgc gctgagcagc agcagcatcg cggccaccag
     1981 ggagagcttc atgctagcgg atctgacggt tcactaaacc agctctgctt atatagacct
     2041 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     2101 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     2161 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     2221 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     2281 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     2341 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     2401 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     2461 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     2521 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     2581 aataactaat gcatggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2641 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2701 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2761 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2821 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2881 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2941 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3001 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3061 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3121 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3181 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3241 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3301 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3361 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3421 aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg
     3481 ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa
     3541 acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     3601 gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt
     3661 attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc
     3721 cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     3781 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     3841 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     3901 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     3961 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     4021 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     4081 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     4141 cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga
     4201 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     4261 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     4321 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     4381 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     4441 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     4501 ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     4561 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     4621 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     4681 cttgttcaat catgcgaaac gatcctcatc ctgtctct
//
