LOCUS       40924_1239        4759 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses GST-HA-GFP11 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4759)
  AUTHORS   Ruan L, Zhou C, Jin E, Kucharavy A, Zhang Y, Wen Z, Florens L, Li R
  TITLE     Cytosolic proteostasis through importing of misfolded proteins into 
            mitochondria.
  JOURNAL   Nature. 2017 Mar 16;543(7645):443-446. doi: 10.1038/nature21695. 
            Epub 2017 Mar 1.
  PUBMED    28241148
REFERENCE   2  (bases 1 to 4759)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4759)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature21695"; journalName: "Nature"; date: "2017-03-16-
            16"; volume: "543"; issue: "7645"; pages: "443-446"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        124..427
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        428..631
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             691..1344
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             1372..1398
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             1438..1485
                     /codon_start=1
                     /label=GFP11
                     /note="11th beta-strand of superfolder GFP"
                     /translation="RDHMVLHEYVNAAGIT"
     polyA_signal    1610..1731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1738..2193)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2220..2324
                     /label=AmpR promoter
     promoter        2326..2683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2718..3509
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3700..3719)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3744..3791
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4120..4708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
       61 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      601 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      661 ctaccggact cagatctcga gctcaagctt atgtccccta tactaggtta ttggaaaatt
      721 aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag
      781 catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg
      841 gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc
      901 atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca
      961 gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca
     1021 tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg
     1081 aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat
     1141 cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg
     1201 gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat
     1261 aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt
     1321 ggtggtggcg accatcctcc aaaaggatcc ggtgacggtc cggtcgccac ctacccatac
     1381 gatgttccag attacgctgg tcgacggatc ggtggtggtt ctggaggcgg ttctatgaga
     1441 gatcatatgg ttttgcatga atatgttaat gctgctggta ttacttaagc ggccgcgact
     1501 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     1561 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1621 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1681 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     1741 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1801 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1861 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1921 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1981 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2041 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2101 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2161 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     2221 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2281 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     2341 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2401 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2461 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2521 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2581 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2641 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     2701 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2761 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2821 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2881 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2941 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3001 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3061 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3121 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3181 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3241 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3301 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3361 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3421 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3481 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3541 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3601 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3661 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     3721 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     3781 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3841 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3901 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     3961 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4021 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4081 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4141 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4201 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4261 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4321 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4381 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4441 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4501 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4561 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4621 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4681 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4741 ggccttttgc tggcctttt
//
