LOCUS       sequence-352621-        7071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER protein alpha-1-antitrypsin containing E342K mutation (ATZ 
            mutant) fused to blue fluorescence protein (BFP) tag protein, an HA 
            tag and the eleventh beta-strand of GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7071)
  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 7071)
  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: pATZ-BFP-HA-GFP11
FEATURES             Location/Qualifiers
     source          1..7071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        66..423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        66..257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(89..109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      274..409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     336..355
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             454..1491
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1470..1478
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    1515..1563
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1719..1736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1890..2478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1970..1989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2678..3469,3470..3538))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    3643..3691
                     /note="synthetic polyadenylation signal"
     misc_feature    3705..3796
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3745..3764
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        3947..4326
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4327..4530
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4481..4501
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4527..4551
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5852..6550
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    6551..6616
                     /label=MCS
                     /note="multiple cloning site"
     CDS             6662..6688
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6728..6775
                     /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"
     polyA_signal    6822..6943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6833..6852)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6887..6906
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ccgctgatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttagct
       61 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      361 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      421 aaaaagctcg attcttctga cactagcgcc accatgaaga agcccgaact caccgctacc
      481 agcgttgaaa aatttctcat cgagaagttc gacagtgtga gcgacctgat gcagttgtcg
      541 gagggcgaag agagccgagc cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg
      601 gtgaatagct gcgctgatgg cttctacaaa gaccgctacg tgtaccgcca cttcgccagc
      661 gctgcactac ccatccccga agtgttggac atcggcgagt tcagcgagag cctgacatac
      721 tgcatcagta gacgcgccca aggcgttact ctccaagacc tccccgaaac agagctgcct
      781 gctgtgttac agcctgtcgc cgaagctatg gatgctattg ccgccgccga cctcagtcaa
      841 accagcggct tcggcccatt cgggccccaa ggcatcggcc agtacacaac ctggcgggat
      901 ttcatttgcg ccattgctga tccccatgtc taccactggc agaccgtgat ggacgacacc
      961 gtgtccgcca gcgtagctca agccctggac gaactgatgc tgtgggccga agactgtccc
     1021 gaggtgcgcc acctcgtcca tgccgacttc ggcagcaaca acgtcctgac cgacaacggc
     1081 cgcatcaccg ccgtaatcga ctggtccgaa gctatgttcg gggacagtca gtacgaggtg
     1141 gccaacatct tcttctggcg gccctggctg gcttgcatgg agcagcagac tcgctacttc
     1201 gagcgccggc atcccgagct ggccggcagc cctcgtctgc gagcctacat gctgcgcatc
     1261 ggcctggatc agctctacca gagcctcgtg gacggcaact tcgacgatgc tgcctgggct
     1321 caaggccgct gcgatgccat cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc
     1381 gctcgccgga gcgcagccgt atggaccgac ggctgcgtcg aggtgctggc cgacagcggc
     1441 aaccgccggc ccagtacacg accgcgcgct aaggaggtag gtcgagttta aactctagaa
     1501 ccggtcatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     1561 gtgttcgaac tagatgctgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     1621 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     1681 aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc
     1741 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1801 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1861 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1921 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1981 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2041 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2101 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2161 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2221 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2281 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2341 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2401 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2461 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2521 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2581 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2641 tctgacagcg gccgcaaatg ctaaaccact gcagtggtta ccagtgcttg atcagtgagg
     2701 caccgatctc agcgatctgc ctatttcgtt cgtccatagt ggcctgactc cccgtcgtgt
     2761 agatcactac gattcgtgag ggcttaccat caggccccag cgcagcaatg atgccgcgag
     2821 agccgcgttc accggccccc gatttgtcag caatgaacca gccagcaggg agggccgagc
     2881 gaagaagtgg tcctgctact ttgtccgcct ccatccagtc tatgagctgc tgtcgtgatg
     2941 ctagagtaag aagttcgcca gtgagtagtt tccgaagagt tgtggccatt gctactggca
     3001 tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa ctctggttcc cagcggtcaa
     3061 gccgggtcac atgatcaccc atattatgaa gaaatgcagt cagctcctta gggcctccga
     3121 tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat ggtaatggca gcactacaca
     3181 attctcttac cgtcatgcca tccgtaagat gcttttccgt gaccggcgag tactcaacca
     3241 agtcgttttg tgagtagtgt atacggcgac caagctgctc ttgcccggcg tctatacggg
     3301 acaacaccgc gccacatagc agtactttga aagtgctcat catcgggaat cgttcttcgg
     3361 ggcggaaaga ctcaaggatc ttgccgctat tgagatccag ttcgatatag cccactcttg
     3421 cacccagttg atcttcagca tcttttactt tcaccagcgt ttcggggtgt gcaaaaacag
     3481 gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg aaaatgttgg atgctcatac
     3541 tcgtcctttt tcaatattat tgaagcattt atcagggtta ctagtacgtc tctcaaggat
     3601 aagtaagtaa tattaaggta cgggaggtat tggacaggcc gcaataaaat atctttattt
     3661 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat
     3721 caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt
     3781 gccagaacat ttctctggcc taactggcct caatattggc cattagccat attattcatt
     3841 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     3901 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttggca ttgattattg
     3961 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     4021 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     4081 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     4141 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     4201 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4261 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4321 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     4381 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4441 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
     4501 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cactagaagc
     4561 ttcgatgccg tcttctgtct cgtggggcat cctcctgctg gcaggcctgt gctgcctggt
     4621 ccctgtctcc ctggctgagg atccccaggg agatgctgcc cagaagacag atacatccca
     4681 ccatgatcag gatcacccaa ccttcaacaa gatcaccccc aacctggctg agttcgcctt
     4741 cagcctatac cgccagctgg cacaccagtc caacagcacc aatatcttct tctccccagt
     4801 gagcatcgct acagcctttg caatgctctc cctggggacc aaggctgaca ctcacgatga
     4861 aatcctggag ggcctgaatt tcaacctcac ggagattccg gaggctcaga tccatgaagg
     4921 cttccaggaa ctcctccgta ccctcaacca gccagacagc cagctccagc tgaccaccgg
     4981 caatggcctg ttcctcagcg agggcctgaa gctagtggat aagtttttgg aggatgttaa
     5041 aaagttgtac cactcagaag ccttcactgt caacttcggg gacaccgaag aggccaagaa
     5101 acagatcaac gattacgtgg agaagggtac tcaagggaaa attgtggatt tggtcaagga
     5161 gcttgacaga gacacagttt ttgctctggt gaattacatc ttctttaaag gcaaatggga
     5221 gagacccttt gaagtcaagg acaccgagga agaggacttc cacgtggacc aggtgaccac
     5281 cgtgaaggtg cctatgatga agcgtttagg catgtttaac atccagcact gtaagaagct
     5341 gtccagctgg gtgctgctga tgaaatacct gggcaatgcc accgccatct tcttcctgcc
     5401 tgatgagggg aaactacagc acctggaaaa tgaactcacc cacgatatca tcaccaagtt
     5461 cctggaaaat gaagacagaa ggtctgccag cttacattta cccaaactgt ccattactgg
     5521 aacctatgat ctgaagagcg tcctgggtca actgggcatc actaaggtct tcagcaatgg
     5581 ggctgacctc tccggggtca cagaggaggc acccctgaag ctctccaagg ccgtgcataa
     5641 ggctgtgctg accatcgaca agaaagggac tgaagctgct ggggccatgt ttttagaggc
     5701 catacccatg tctatccccc ccgaggtcaa gttcaacaaa ccctttgtct tcttaatgat
     5761 tgaacaaaat accaagtctc ccctcttcat gggaaaagtg gtgaatccca cccaaaaacc
     5821 cgggggatcg ggtgacggtc cggtcgccac catgagcgag ctgattaagg agaacatgca
     5881 catgaagctg tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg
     5941 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc
     6001 tctccccttc gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat
     6061 caaccacacc cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg
     6121 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
     6181 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg
     6241 ccctgtgatg cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc
     6301 tgacggcggc ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct
     6361 gatcgcaaac atcaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc
     6421 tggcgtctac tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta
     6481 cgtcgagcag cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca
     6541 caagcttaat tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac
     6601 cgcgggcccg ggatccaccg gatctagacc cgggggatcg ggtgacggtc cggtcgccac
     6661 ctacccatac gatgttccag attacgctgg tcgacggatc ggtggtggtt ctggaggcgg
     6721 ttctatgaga gatcatatgg ttttgcatga atatgttaat gctgctggta ttacttaata
     6781 attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     6841 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     6901 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     6961 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     7021 ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat tgggcgctct t
//
