LOCUS       sequence-304841-        7653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV:EGFP (as control for pXG237).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7653)
  AUTHORS   Guo X, Aviles G, Liu Y, Tian R, Unger BA, Lin YT, Wiita AP, Xu K, 
            Correia MA, Kampmann M
  TITLE     Mitochondrial stress is relayed to the cytosol by an OMA1-DELE1-HRI 
            pathway.
  JOURNAL   Nature. 2020 Mar;579(7799):427-432. doi: 10.1038/s41586-020-2078-2. 
            Epub 2020 Mar 4.
  PUBMED    32132707
REFERENCE   2  (bases 1 to 7653)
  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"; doi: 
            "10.1038/s41586-020-2078-2"; journalName: "Nature"; date: "2020-03";
            volume: "579"; issue: "7799"; pages: "427-432"
COMMENT     Sequence Label: pXG260
FEATURES             Location/Qualifiers
     source          1..7653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(72..176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(251..270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        442..821
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        823..1021
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             1039..1219
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1266..1391
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1888..2121
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2306..2350
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2648..2765
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(3018..3051)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     3055..3075
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     enhancer        3268..3571
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3572..3775
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3904..3913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3910..3912,3913..3915,3916..4629)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3910..3912,3913..3915,3916..4626)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3955..3976)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4216..4235)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4563..4584
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4657..4690)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4746..5334
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4799..4819)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5217..5228)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5337..5353)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5338..5354)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5863..6043
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6105..6693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6185..6204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6864..7653,1..2,3..71))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7487..7506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
       61 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      121 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      181 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
      241 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
      301 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      361 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      421 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      481 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      541 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      601 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      661 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      721 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      781 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      841 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      901 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      961 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     1021 gcgcgttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     1081 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     1141 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     1201 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     1261 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     1321 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     1381 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     1441 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     1501 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     1561 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     1621 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     1681 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccggccgct
     1741 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     1801 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     1861 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     1921 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     1981 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     2041 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     2101 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     2161 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     2221 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     2281 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     2341 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     2401 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     2461 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     2521 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     2581 tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca
     2641 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     2701 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     2761 attttcgggt ttattacagg gacagcagag atccagtttg gttagtaccg ggcccgctct
     2821 agagatccga cgcgccatct ctaggcccgc gccggccccc tcgcacagac ttgtgggaga
     2881 agctcggcta ctcccctgcc ccggttaatt tgcatataat atttcctagt aactatagag
     2941 gcttaatgtg cgataaaaga cagataatct gttcttttta atactagcta cattttacat
     3001 gataggcttg gatttctata acttcgtata gcatacatta tacgaagtta taaacagcac
     3061 aaaaggaaac tcaccctaac tgtaaagtaa ttgtgtgttt tgagactata aatatccctt
     3121 ggagaaaagc cttgttaacg cgcggtgacc ctcgagtact aggatccatt aggcggccgc
     3181 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     3241 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     3301 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3361 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3421 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3481 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3541 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     3601 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     3661 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     3721 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     3781 agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagtttct actttgcccg
     3841 cccacagatg tagttttctc tgcgcgtgtg cgttttccct cctccccgcc ctcagggtcc
     3901 acggccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3961 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4021 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4081 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4141 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4201 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4261 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4321 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4381 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4441 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4501 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4561 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4621 tacaagtagg cctaggcgaa ttcgtcgagg gacctaataa cttcgtatag catacattat
     4681 acgaagttat acatgtttaa gggttccggt tccactaggt acaattcgat atcaagctta
     4741 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4801 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4861 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4921 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4981 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5041 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5101 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     5161 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5221 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5281 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     5341 accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     5401 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     5461 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5521 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
     5581 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
     5641 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     5701 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
     5761 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
     5821 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
     5881 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     5941 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6001 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc
     6061 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6301 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6541 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6601 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6661 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6721 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6781 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6841 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6901 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6961 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7021 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7081 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7141 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7201 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7261 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7321 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7381 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7441 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7501 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7561 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7621 tcagcatctt ttactttcac cagcgtttct ggg
//
