LOCUS       sequence-352608-        7580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER protein collagen beta (1-O) galactosyltransferase fused to eGFP 
            at the C-terminus, myc tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7580)
  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 7580)
  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-eGFP-KDEL
FEATURES             Location/Qualifiers
     source          1..7580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        120..477
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        120..311
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(143..163)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      328..463
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     390..409
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             508..1545
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1524..1532
                     /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    1569..1617
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1773..1790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1944..2532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2024..2043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2732..3523,3524..3592))
                     /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    3697..3745
                     /note="synthetic polyadenylation signal"
     misc_feature    3759..3850
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3799..3818
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        4001..4380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4381..4584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4535..4555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4581..4605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4646..4655
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5333..5344)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             6535..6564
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(6601..6603,6604..6606,6607..7317)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6601..6603,6604..6606,6607..7317)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6646..6667)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6907..6926)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7254..7275
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7385..7506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7396..7415)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7450..7469
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cctctacaaa tgtggtaaaa tcgataagga tccgtttgcg tattgggcgc tcttccgctg
       61 atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt agctaccttc
      121 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
      181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
      241 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      301 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
      361 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
      421 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
      481 ctcgattctt ctgacactag cgccaccatg aagaagcccg aactcaccgc taccagcgtt
      541 gaaaaatttc tcatcgagaa gttcgacagt gtgagcgacc tgatgcagtt gtcggagggc
      601 gaagagagcc gagccttcag cttcgatgtc ggcggacgcg gctatgtact gcgggtgaat
      661 agctgcgctg atggcttcta caaagaccgc tacgtgtacc gccacttcgc cagcgctgca
      721 ctacccatcc ccgaagtgtt ggacatcggc gagttcagcg agagcctgac atactgcatc
      781 agtagacgcg cccaaggcgt tactctccaa gacctccccg aaacagagct gcctgctgtg
      841 ttacagcctg tcgccgaagc tatggatgct attgccgccg ccgacctcag tcaaaccagc
      901 ggcttcggcc cattcgggcc ccaaggcatc ggccagtaca caacctggcg ggatttcatt
      961 tgcgccattg ctgatcccca tgtctaccac tggcagaccg tgatggacga caccgtgtcc
     1021 gccagcgtag ctcaagccct ggacgaactg atgctgtggg ccgaagactg tcccgaggtg
     1081 cgccacctcg tccatgccga cttcggcagc aacaacgtcc tgaccgacaa cggccgcatc
     1141 accgccgtaa tcgactggtc cgaagctatg ttcggggaca gtcagtacga ggtggccaac
     1201 atcttcttct ggcggccctg gctggcttgc atggagcagc agactcgcta cttcgagcgc
     1261 cggcatcccg agctggccgg cagccctcgt ctgcgagcct acatgctgcg catcggcctg
     1321 gatcagctct accagagcct cgtggacggc aacttcgacg atgctgcctg ggctcaaggc
     1381 cgctgcgatg ccatcgtccg cagcggggcc ggcaccgtcg gtcgcacaca aatcgctcgc
     1441 cggagcgcag ccgtatggac cgacggctgc gtcgaggtgc tggccgacag cggcaaccgc
     1501 cggcccagta cacgaccgcg cgctaaggag gtaggtcgag tttaaactct agaaccggtc
     1561 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgttc
     1621 gaactagatg ctgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     1681 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     1741 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1801 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1861 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1921 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1981 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2041 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2101 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2161 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2221 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2281 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2341 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2401 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2461 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2521 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2581 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2641 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2701 agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga
     2761 tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca
     2821 ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc
     2881 gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa
     2941 gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag
     3001 taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg
     3061 tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg
     3121 tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg
     3181 tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc
     3241 ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt
     3301 tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca
     3361 ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga
     3421 aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca
     3481 gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc
     3541 aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc
     3601 tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa
     3661 gtaatattaa ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta
     3721 catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac
     3781 aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga
     3841 acatttctct ggcctaactg gcctcaatat tggccattag ccatattatt cattggttat
     3901 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     3961 tacatttata ttggctcatg tccaatatga ccgccatgtt ggcattgatt attgactagt
     4021 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4081 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4141 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4201 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4261 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     4321 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4381 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
     4441 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     4501 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
     4561 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcactag aagcttggta
     4621 ccgaggagat ctgccgccgc gatcgccatg gcggcggccc cacgcgcggg ccggcggcgc
     4681 gggcagccgc tcctggcgct gctgcttctg ctgctggcgc cactgccgcc gggggccccg
     4741 ccgggcgccg acgcctactt tcccgaggag cgctggagcc cggagtcgcc cctgcaggcg
     4801 ccgcgcgtgc tcatcgcgct gttggcgcga aacgcggccc acgcgttgcc caccacgctg
     4861 ggcgcactcg agcggctgcg gcacccgcgg gagcgcacgg cgctatgggt ggctacggac
     4921 cacaacatgg ataacacgtc aactgtgctg cgggagtggc tggtggccgt gaagagtttg
     4981 taccattccg tggagtggcg gccagcagag gagcccaggt cctacccgga cgaggaaggc
     5041 ccgaaacact ggtctgactc acgctacgag catgtcatga agttgcgcca ggcagccctg
     5101 aaatcagctc gagacatgtg ggctgattac atcctgtttg tagatgcgga caacctgatc
     5161 ctcaaccctg acacactgag cctgctcatc gctgagaaca agacggtggt cgcccccatg
     5221 ctggattccc gggctgcgta ctccaacttc tggtgtggaa tgacttccca gggctactac
     5281 aagcgcacac ctgcctacat ccctatccgc aagcgagacc gccggggctg ctttgcagtt
     5341 cccatggtgc actcgacctt cctgatcgac ctgcggaagg cggcgtccag gaacctggcc
     5401 ttctacccac ctcaccctga ctacacctgg tcctttgacg acatcatcgt ctttgccttc
     5461 tcctgcaagc aggcagaggt tcagatgtat gtgtgcaaca aggaggagta cggattcttg
     5521 ccagtgccat tgcgcgccca cagcaccctc caggatgagg ccgagagctt catgcatgtg
     5581 cagctggagg tcatggtgaa gcacccgccc gcagagccct cccgcttcat ctcggctccc
     5641 accaagacac cggacaagat gggcttcgac gaggtcttca tgatcaacct gaggcggcgg
     5701 caggaccggc gggagcgcat gctgcgggcg ctgcaggcac aggagatcga gtgccggctg
     5761 gtggaggccg tggacggcaa agccatgaac accagccagg tggaggcgct ggggatccag
     5821 atgctgcctg gctaccggga cccctaccac ggccggcccc tcaccaaggg tgagctgggc
     5881 tgcttcctga gccactacaa catctggaag gaggtggtgg accgggggct gcagaaatcg
     5941 cttgtgtttg aggatgacct gcgttttgag atcttcttca agagacgtct gatgaacctc
     6001 atgcgggatg tggagcggga gggcctggac tgggacctca tctatgtggg ccggaagcgg
     6061 atgcaggtgg agcaccccga gaaggctgtg cctcgcgtga ggaacctggt ggaggccgac
     6121 tattcctact ggaccctggc ctacgtgatc tccctgcaag gcgcccgcaa actgctggct
     6181 gctgagccgc tctccaagat gctgcctgtg gacgagttcc tgcccgtcat gttcgacaaa
     6241 cacccagtgt ccgagtacaa ggcccacttc tccctccgca acctgcatgc cttctctgtg
     6301 gagccgctgc tcatctaccc cacacactac acaggagacg atggctatgt gagtgacacc
     6361 gagacctcag tcgtatggaa caatgagcac gtcaagaccg actgggaccg cgccaagtcc
     6421 cagaagatgc gggagcagca ggcactgagc cgtgaggcca agaactcgga cgtgctccag
     6481 tccccactgg acagtgctgc ccgggatgaa ctcagcacgc gtacgcggcc gctcgagcag
     6541 aaactcatct cagaagagga tctgatcggt ccgatcggtg gtggttctgg aggcggttct
     6601 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     6661 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     6721 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     6781 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     6841 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     6901 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     6961 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     7021 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     7081 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     7141 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     7201 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     7261 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagaaa
     7321 aagaccaaag atgaactcta ataattctag agtcggggcg gccggccgct tcgagcagac
     7381 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     7441 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     7501 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     7561 gttttttaaa gcaagtaaaa
//
