LOCUS       sequence-352637-        5388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Eleventh beta-strand of GFP (GFP11) fused to a mitochondria signal 
            sequence for mitochondrial targeting, HA tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5388)
  AUTHORS   Cortes Sanchon A, Santhosh Kumar H, Mantovani M, Osinnii I, Mateos 
            JM, Kaech A, Shcherbakov D, Akbergenov R, Bottger EC
  TITLE     ER-misfolded proteins become sequestered with mitochondria and 
            impair mitochondrial function.
  JOURNAL   Commun Biol. 2021 Dec 2;4(1):1350. doi: 10.1038/s42003-021-02873-w.
  PUBMED    34857875
REFERENCE   2  (bases 1 to 5388)
  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: "Commun 
            Biol."; date: "2021-12-2"; volume: "4(1):1350. doi"; pages: " 
            10.1038/s42003-021-02873-w"
COMMENT     Sequence Label: pMTS-HA-GFP11
FEATURES             Location/Qualifiers
     source          1..5388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        166..357
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     314..334
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(359..463)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      490..945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(577..596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     787..808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    952..1073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(963..982)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1198..1245)
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             complement(1285..1311)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1622..1825)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1651..1671)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1826..2129
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2304..2892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2384..2403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3078..3869,3870..3938))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3701..3720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3939..4043)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4111..4129
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    4157..4238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4355..5380)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(4355..5380)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(join(5388,1..357))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
       61 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      121 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      181 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      241 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      301 ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac
      361 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
      421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      481 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
      541 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
      601 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
      661 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
      721 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
      781 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
      841 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
      901 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacgcc ttaagataca
      961 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1021 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1081 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1141 agtaaaacct ctacaaatgt ggtatggctg attatgatct agagtcgcgg ccgcttaagt
     1201 aataccagca gcattaacat attcatgcaa aaccatatga tctctcatag aaccgcctcc
     1261 agaaccacca ccgatccgtc gaccagcgta atctggaaca tcgtatgggt aggtggcgac
     1321 cggaccgtca ccggatccgg aagagtaggc gcgcttctgg aaagcctggc gggtggtggc
     1381 gttgacgatg gaggtcatct gggtgcgctt gagggtctgg aggggggagc cagtctggat
     1441 ggtgcgcttg ctgacctgag caacgcggac agcagggcgg gcaaccttgg cggaagcagc
     1501 catccgggag gccaggcgag aggcgaggac acgagtggag gccatgtcga ctgcagaatt
     1561 cgaagcttga gctcgagatc tgagtccggt agcgctagcg gatctgacgg ttcactaaac
     1621 cagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg
     1681 cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat
     1741 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
     1801 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
     1861 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     1921 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     1981 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     2041 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     2101 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     2161 accccgtaat tgattactat taataactaa tgcatggcgg taatacggtt atccacagaa
     2221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2401 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2461 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2521 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2581 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2641 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2701 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2761 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2821 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg
     3061 gcagggcctg gcgcgcctta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3121 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3361 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3661 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     3721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4081 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgccat
     4141 ggttgtggat cgcgaggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4201 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     4261 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     4321 aatgtggtat ggctgattat gatcctctag aactctattc ctttgccctc ggacgagtgc
     4381 tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg
     4441 cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt
     4501 cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga
     4561 gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct gcaagctccg
     4621 gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga
     4681 agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga
     4741 ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga
     4801 ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga
     4861 cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa
     4921 tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc
     4981 cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatggcc tccgcgaccg
     5041 gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac
     5101 ggcgggagat gcaataggtc aggctctcgc tgaattcccc aatgtcaagc acttccggaa
     5161 tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg
     5221 cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag
     5281 attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa
     5341 acttctcgac agacgtcgcg gtgagttcag gctttttcat atcgatct
//
