LOCUS       sequence-352642-        6261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER protein alpha-1-antitrypsin containing E342K mutation (ATZ 
            mutant) fused to 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 6261)
  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 6261)
  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-HA-GFP11
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        92..449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        92..283
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(115..135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      300..435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     362..381
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             480..1517
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1496..1504
                     /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    1541..1589
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1745..1762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1916..2504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1996..2015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2704..3495,3496..3564))
                     /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    3669..3717
                     /note="synthetic polyadenylation signal"
     misc_feature    3731..3822
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3771..3790
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        3973..4352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4353..4556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4507..4527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4553..4577
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5878..5904
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5944..5991
                     /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    6038..6159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6049..6068)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6103..6122
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gatccgtttg cgtattgggc gctcttccgc tgatctgcgc agcaccatgg cctgaaataa
       61 cctctgaaag aggaacttgg ttagctacct tctgaggcgg aaagaaccag ctgtggaatg
      121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
      241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
      301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
      361 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
      421 gcttttttgg aggcctaggc ttttgcaaaa agctcgattc ttctgacact agcgccacca
      481 tgaagaagcc cgaactcacc gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca
      541 gtgtgagcga cctgatgcag ttgtcggagg gcgaagagag ccgagccttc agcttcgatg
      601 tcggcggacg cggctatgta ctgcgggtga atagctgcgc tgatggcttc tacaaagacc
      661 gctacgtgta ccgccacttc gccagcgctg cactacccat ccccgaagtg ttggacatcg
      721 gcgagttcag cgagagcctg acatactgca tcagtagacg cgcccaaggc gttactctcc
      781 aagacctccc cgaaacagag ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg
      841 ctattgccgc cgccgacctc agtcaaacca gcggcttcgg cccattcggg ccccaaggca
      901 tcggccagta cacaacctgg cgggatttca tttgcgccat tgctgatccc catgtctacc
      961 actggcagac cgtgatggac gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac
     1021 tgatgctgtg ggccgaagac tgtcccgagg tgcgccacct cgtccatgcc gacttcggca
     1081 gcaacaacgt cctgaccgac aacggccgca tcaccgccgt aatcgactgg tccgaagcta
     1141 tgttcgggga cagtcagtac gaggtggcca acatcttctt ctggcggccc tggctggctt
     1201 gcatggagca gcagactcgc tacttcgagc gccggcatcc cgagctggcc ggcagccctc
     1261 gtctgcgagc ctacatgctg cgcatcggcc tggatcagct ctaccagagc ctcgtggacg
     1321 gcaacttcga cgatgctgcc tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg
     1381 ccggcaccgt cggtcgcaca caaatcgctc gccggagcgc agccgtatgg accgacggct
     1441 gcgtcgaggt gctggccgac agcggcaacc gccggcccag tacacgaccg cgcgctaagg
     1501 aggtaggtcg agtttaaact ctagaaccgg tcatggccgc aataaaatat ctttattttc
     1561 attacatctg tgtgttggtt ttttgtgtgt tcgaactaga tgctgtcgac cgatgccctt
     1621 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     1681 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg
     1741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2341 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2641 tctaaagtat atatgagtaa acttggtctg acagcggccg caaatgctaa accactgcag
     2701 tggttaccag tgcttgatca gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc
     2761 catagtggcc tgactccccg tcgtgtagat cactacgatt cgtgagggct taccatcagg
     2821 ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat
     2881 gaaccagcca gcagggaggg ccgagcgaag aagtggtcct gctactttgt ccgcctccat
     2941 ccagtctatg agctgctgtc gtgatgctag agtaagaagt tcgccagtga gtagtttccg
     3001 aagagttgtg gccattgcta ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc
     3061 gttcaactct ggttcccagc ggtcaagccg ggtcacatga tcacccatat tatgaagaaa
     3121 tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc
     3181 gctcatggta atggcagcac tacacaattc tcttaccgtc atgccatccg taagatgctt
     3241 ttccgtgacc ggcgagtact caaccaagtc gttttgtgag tagtgtatac ggcgaccaag
     3301 ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca catagcagta ctttgaaagt
     3361 gctcatcatc gggaatcgtt cttcggggcg gaaagactca aggatcttgc cgctattgag
     3421 atccagttcg atatagccca ctcttgcacc cagttgatct tcagcatctt ttactttcac
     3481 cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc
     3541 gacacgaaaa tgttggatgc tcatactcgt cctttttcaa tattattgaa gcatttatca
     3601 gggttactag tacgtctctc aaggataagt aagtaatatt aaggtacggg aggtattgga
     3661 caggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     3721 cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat
     3781 aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctggcctaac tggcctcaat
     3841 attggccatt agccatatta ttcattggtt atatagcata aatcaatatt ggctattggc
     3901 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     3961 gaccgccatg ttggcattga ttattgacta gttattaata gtaatcaatt acggggtcat
     4021 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     4081 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4141 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4201 tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta
     4261 aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt
     4321 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg
     4381 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     4441 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc
     4501 cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     4561 tagtgaaccg tcagatcact agaagcttcg atgccgtctt ctgtctcgtg gggcatcctc
     4621 ctgctggcag gcctgtgctg cctggtccct gtctccctgg ctgaggatcc ccagggagat
     4681 gctgcccaga agacagatac atcccaccat gatcaggatc acccaacctt caacaagatc
     4741 acccccaacc tggctgagtt cgccttcagc ctataccgcc agctggcaca ccagtccaac
     4801 agcaccaata tcttcttctc cccagtgagc atcgctacag cctttgcaat gctctccctg
     4861 gggaccaagg ctgacactca cgatgaaatc ctggagggcc tgaatttcaa cctcacggag
     4921 attccggagg ctcagatcca tgaaggcttc caggaactcc tccgtaccct caaccagcca
     4981 gacagccagc tccagctgac caccggcaat ggcctgttcc tcagcgaggg cctgaagcta
     5041 gtggataagt ttttggagga tgttaaaaag ttgtaccact cagaagcctt cactgtcaac
     5101 ttcggggaca ccgaagaggc caagaaacag atcaacgatt acgtggagaa gggtactcaa
     5161 gggaaaattg tggatttggt caaggagctt gacagagaca cagtttttgc tctggtgaat
     5221 tacatcttct ttaaaggcaa atgggagaga ccctttgaag tcaaggacac cgaggaagag
     5281 gacttccacg tggaccaggt gaccaccgtg aaggtgccta tgatgaagcg tttaggcatg
     5341 tttaacatcc agcactgtaa gaagctgtcc agctgggtgc tgctgatgaa atacctgggc
     5401 aatgccaccg ccatcttctt cctgcctgat gaggggaaac tacagcacct ggaaaatgaa
     5461 ctcacccacg atatcatcac caagttcctg gaaaatgaag acagaaggtc tgccagctta
     5521 catttaccca aactgtccat tactggaacc tatgatctga agagcgtcct gggtcaactg
     5581 ggcatcacta aggtcttcag caatggggct gacctctccg gggtcacaga ggaggcaccc
     5641 ctgaagctct ccaaggccgt gcataaggct gtgctgacca tcgacaagaa agggactgaa
     5701 gctgctgggg ccatgttttt agaggccata cccatgtcta tcccccccga ggtcaagttc
     5761 aacaaaccct ttgtcttctt aatgattgaa caaaatacca agtctcccct cttcatggga
     5821 aaagtggtga atcccaccca aaaacccggg ggatcgggtg acggtccggt cgccacctac
     5881 ccatacgatg ttccagatta cgctggtcga cggatcggtg gtggttctgg aggcggttct
     5941 atgagagatc atatggtttt gcatgaatat gttaatgctg ctggtattac ttaataattc
     6001 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     6061 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     6121 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     6181 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     6241 aaatgtggta aaatcgataa g
//
