LOCUS       sequence-207851-        5229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For recruitment of ER.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5229)
  AUTHORS   Zewe JP, Wills RC, Sangappa S, Goulden BD, Hammond GR
  TITLE     SAC1 degrades its lipid substrate PtdIns4P in the endoplasmic 
            reticulum to maintain a steep chemical gradient with donor 
            membranes.
  JOURNAL   Elife. 2018 Feb 20;7. pii: 35588. doi: 10.7554/eLife.35588.
  PUBMED    29461204
REFERENCE   2  (bases 1 to 5229)
  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: "Elife."; 
            date: "2018-02-20"; volume: "7. pii: 35588. doi"; pages: " 
            10.7554/eLife.35588"
COMMENT     Sequence Label: mTagBFP2-FKBP-CYB5Atail
FEATURES             Location/Qualifiers
     source          1..5229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             612..1313
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             1374..1694
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    2016..2137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2053..2072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2107..2126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2144..2599)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2281..2302)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2493..2512
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2626..2730
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2732..3089
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2732..2923
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2755..2775)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2940..3075
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3124..3918
                     /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             3124..3918
                     /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(3178..3197)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3788..3807
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4106..4125)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4150..4197
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4526..5114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5015..5034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      601 cggtcgccac catggtgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg
      661 agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg
      721 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
      781 acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca
      841 tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
      901 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
      961 tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga
     1021 aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag
     1081 gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca aacgccaaga
     1141 ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg
     1201 actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg
     1261 tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt aattccggac
     1321 tcagatctcg aagcgcggcc gcgggagcag gaggagcagc tcgagcggcg ctgggagtgc
     1381 aggtggaaac catctcccca ggagacgggc gcaccttccc caagcgcggc cagacctgcg
     1441 tggtgcacta caccgggatg cttgaagatg gaaagaaatt tgattcctcc cgggacagaa
     1501 acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg
     1561 ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg
     1621 gtgccactgg gcacccaggc atcatcccac cacatgccac tctcgtcttc gatgtggagc
     1681 ttctaaaact ggaaagtgct ggtggtagtg ctggtggtag tgctggtggt agtgctggtg
     1741 gtagtgctgg tggtccgcga gctcaagctt cgaattctat cactactatt gattctagtt
     1801 ccagttggtg gaccaactgg gtgatccctg ccatctctgc agtggccgtc gccttgatgt
     1861 atcgcctata catggcagag gactgaggat ccaccggatc tagataactg atcataatca
     1921 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     1981 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2041 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2101 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     2161 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2221 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2281 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2341 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2401 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2461 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2521 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2581 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2641 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2701 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     2761 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2881 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3001 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3061 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3121 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3181 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3241 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3301 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3361 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3421 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3481 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3541 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3601 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3661 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3721 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3781 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3841 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3901 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3961 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4021 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4081 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4141 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4201 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4261 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4321 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4381 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4441 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5161 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5221 ccatgcatt
//
