LOCUS       sequence-352616-        6959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER protein collagen beta (1-O) galactosyltransferase with a 
            C-terminal Myc tag, 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 6959)
  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 6959)
  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: pGal-Trans-Myc-HA-GFP11-KDEL
FEATURES             Location/Qualifiers
     source          1..6959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             394..402
                     /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    439..487
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(643..660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(814..1402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(894..913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1602..2393,2394..2462))
                     /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    2567..2615
                     /note="synthetic polyadenylation signal"
     misc_feature    2629..2720
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2669..2688
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        2871..3250
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3251..3454
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3405..3425
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3451..3475
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3516..3525
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4203..4214)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5405..5434
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5453..5479
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5519..5566
                     /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    5634..5755
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5645..5664)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5699..5718
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5949..6306
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5949..6140
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5972..5992)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6157..6292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6219..6238
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(6337..6959,1..415)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
ORIGIN
        1 cggccgcatc accgccgtaa tcgactggtc cgaagctatg ttcggggaca gtcagtacga
       61 ggtggccaac atcttcttct ggcggccctg gctggcttgc atggagcagc agactcgcta
      121 cttcgagcgc cggcatcccg agctggccgg cagccctcgt ctgcgagcct acatgctgcg
      181 catcggcctg gatcagctct accagagcct cgtggacggc aacttcgacg atgctgcctg
      241 ggctcaaggc cgctgcgatg ccatcgtccg cagcggggcc ggcaccgtcg gtcgcacaca
      301 aatcgctcgc cggagcgcag ccgtatggac cgacggctgc gtcgaggtgc tggccgacag
      361 cggcaaccgc cggcccagta cacgaccgcg cgctaaggag gtaggtcgag tttaaactct
      421 agaaccggtc atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
      481 ttgtgtgttc gaactagatg ctgtcgaccg atgcccttga gagccttcaa cccagtcagc
      541 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
      601 atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct
      661 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      721 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      781 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      841 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      901 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      961 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1021 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1081 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1141 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1201 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1261 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1321 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1381 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1441 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1501 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1561 ttggtctgac agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt
     1621 gaggcaccga tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc
     1681 gtgtagatca ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg
     1741 cgagagccgc gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc
     1801 gagcgaagaa gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt
     1861 gatgctagag taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact
     1921 ggcatcgtgg tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg
     1981 tcaagccggg tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct
     2041 ccgatcgttg tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta
     2101 cacaattctc ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca
     2161 accaagtcgt tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata
     2221 cgggacaaca ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct
     2281 tcggggcgga aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact
     2341 cttgcaccca gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa
     2401 acaggcaagc aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc
     2461 atactcgtcc tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa
     2521 ggataagtaa gtaatattaa ggtacgggag gtattggaca ggccgcaata aaatatcttt
     2581 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
     2641 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
     2701 aggtgccaga acatttctct ggcctaactg gcctcaatat tggccattag ccatattatt
     2761 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata
     2821 tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt ggcattgatt
     2881 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2941 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3001 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3061 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3121 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3181 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3241 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc
     3301 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     3361 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag
     3421 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcactag
     3481 aagcttggta ccgaggagat ctgccgccgc gatcgccatg gcggcggccc cacgcgcggg
     3541 ccggcggcgc gggcagccgc tcctggcgct gctgcttctg ctgctggcgc cactgccgcc
     3601 gggggccccg ccgggcgccg acgcctactt tcccgaggag cgctggagcc cggagtcgcc
     3661 cctgcaggcg ccgcgcgtgc tcatcgcgct gttggcgcga aacgcggccc acgcgttgcc
     3721 caccacgctg ggcgcactcg agcggctgcg gcacccgcgg gagcgcacgg cgctatgggt
     3781 ggctacggac cacaacatgg ataacacgtc aactgtgctg cgggagtggc tggtggccgt
     3841 gaagagtttg taccattccg tggagtggcg gccagcagag gagcccaggt cctacccgga
     3901 cgaggaaggc ccgaaacact ggtctgactc acgctacgag catgtcatga agttgcgcca
     3961 ggcagccctg aaatcagctc gagacatgtg ggctgattac atcctgtttg tagatgcgga
     4021 caacctgatc ctcaaccctg acacactgag cctgctcatc gctgagaaca agacggtggt
     4081 cgcccccatg ctggattccc gggctgcgta ctccaacttc tggtgtggaa tgacttccca
     4141 gggctactac aagcgcacac ctgcctacat ccctatccgc aagcgagacc gccggggctg
     4201 ctttgcagtt cccatggtgc actcgacctt cctgatcgac ctgcggaagg cggcgtccag
     4261 gaacctggcc ttctacccac ctcaccctga ctacacctgg tcctttgacg acatcatcgt
     4321 ctttgccttc tcctgcaagc aggcagaggt tcagatgtat gtgtgcaaca aggaggagta
     4381 cggattcttg ccagtgccat tgcgcgccca cagcaccctc caggatgagg ccgagagctt
     4441 catgcatgtg cagctggagg tcatggtgaa gcacccgccc gcagagccct cccgcttcat
     4501 ctcggctccc accaagacac cggacaagat gggcttcgac gaggtcttca tgatcaacct
     4561 gaggcggcgg caggaccggc gggagcgcat gctgcgggcg ctgcaggcac aggagatcga
     4621 gtgccggctg gtggaggccg tggacggcaa agccatgaac accagccagg tggaggcgct
     4681 ggggatccag atgctgcctg gctaccggga cccctaccac ggccggcccc tcaccaaggg
     4741 tgagctgggc tgcttcctga gccactacaa catctggaag gaggtggtgg accgggggct
     4801 gcagaaatcg cttgtgtttg aggatgacct gcgttttgag atcttcttca agagacgtct
     4861 gatgaacctc atgcgggatg tggagcggga gggcctggac tgggacctca tctatgtggg
     4921 ccggaagcgg atgcaggtgg agcaccccga gaaggctgtg cctcgcgtga ggaacctggt
     4981 ggaggccgac tattcctact ggaccctggc ctacgtgatc tccctgcaag gcgcccgcaa
     5041 actgctggct gctgagccgc tctccaagat gctgcctgtg gacgagttcc tgcccgtcat
     5101 gttcgacaaa cacccagtgt ccgagtacaa ggcccacttc tccctccgca acctgcatgc
     5161 cttctctgtg gagccgctgc tcatctaccc cacacactac acaggagacg atggctatgt
     5221 gagtgacacc gagacctcag tcgtatggaa caatgagcac gtcaagaccg actgggaccg
     5281 cgccaagtcc cagaagatgc gggagcagca ggcactgagc cgtgaggcca agaactcgga
     5341 cgtgctccag tccccactgg acagtgctgc ccgggatgaa ctcagcacgc gtacgcggcc
     5401 gctcgagcag aaactcatct cagaagagga tctgatcggt ccggtcgcca cctacccata
     5461 cgatgttcca gattacgctg gtcgacggat cggtggtggt tctggaggcg gttctatgag
     5521 agatcatatg gttttgcatg aatatgttaa tgctgctggt attactaaaa agaccaaaga
     5581 tgaactctaa taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata
     5641 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     5701 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     5761 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     5821 caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtttgcgt attgggcgct
     5881 cttccgctga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     5941 gctaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     6001 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6061 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6121 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6181 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6241 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     6301 tgcaaaaagc tcgattcttc tgacactagc gccaccatga agaagcccga actcaccgct
     6361 accagcgttg aaaaatttct catcgagaag ttcgacagtg tgagcgacct gatgcagttg
     6421 tcggagggcg aagagagccg agccttcagc ttcgatgtcg gcggacgcgg ctatgtactg
     6481 cgggtgaata gctgcgctga tggcttctac aaagaccgct acgtgtaccg ccacttcgcc
     6541 agcgctgcac tacccatccc cgaagtgttg gacatcggcg agttcagcga gagcctgaca
     6601 tactgcatca gtagacgcgc ccaaggcgtt actctccaag acctccccga aacagagctg
     6661 cctgctgtgt tacagcctgt cgccgaagct atggatgcta ttgccgccgc cgacctcagt
     6721 caaaccagcg gcttcggccc attcgggccc caaggcatcg gccagtacac aacctggcgg
     6781 gatttcattt gcgccattgc tgatccccat gtctaccact ggcagaccgt gatggacgac
     6841 accgtgtccg ccagcgtagc tcaagccctg gacgaactga tgctgtgggc cgaagactgt
     6901 cccgaggtgc gccacctcgt ccatgccgac ttcggcagca acaacgtcct gaccgacaa
//
