LOCUS       sequence-458075-        7371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MET 5'UTR upstream of mGFP (uORF1 and uORF2 mutated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7371)
  AUTHORS   Cerqua M, Foiani M, Boccaccio C, Comoglio PM, Altintas DM
  TITLE     The integrated stress response drives MET oncogene overexpression in
            cancers.
  JOURNAL   EMBO J. 2025 Jan 7. doi: 10.1038/s44318-024-00338-4.
  PUBMED    39774381
REFERENCE   2  (bases 1 to 7371)
  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: "EMBO J. 
            2025 Jan 7. doi: 10.1038/s44318-024-00338-4."
COMMENT     Sequence Label: pLenti-uORF1,2-MET
FEATURES             Location/Qualifiers
     source          1..7371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1683..1916
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2101..2145
                     /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    2443..2560
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2643..3022
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3023..3226
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3252..3271
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3252..3270
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3686..4399
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
                     /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK
                     FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG
                     KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA
                     NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE
                     SFSACCHTHGMDELYR"
     protein_bind    complement(4466..4499)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4555..5143
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4608..4628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5026..5037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5146..5162)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5147..5163)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5672..5852
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5914..6502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5994..6013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6579..7238)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7153..7172
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7239..7343)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gctctagaca tgtccaatat gaccgccatg
     2641 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     2701 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2761 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2821 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2881 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     2941 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     3001 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     3061 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     3121 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     3181 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     3241 tcagaatttt gtaatacgac tcactatagg gcggccggga attcagacac gtgctggggc
     3301 gggcaggcga gcgcctcagt ctggtcgcct ggcggtgcct ccggccccaa cgcgcccggg
     3361 ccgccgcggg ccgcgcgcgc cgaagcccgg ctgagtcact ggcagggcag cgcgcgtgtg
     3421 ggaaggggcg gagggagtgc ggccggcggg cgggcggggc gctgggctca gccgggccgc
     3481 aggtgacccg gaggccctcg ccgcccgcgg cgccccgagc gcttgtgagc agttccggag
     3541 ccgagtggag ggcgcgagcc agatgcgggg cgacagctga cttgctgaga ggaggcgggg
     3601 aggcgcggag cgcgcgtgtg gtccttgcgc cgctgacttc tccactggtt cctgggcacc
     3661 gaaagataaa cctctcatac tcgagatgag cgggggcgag gagctgttcg ccggcatcgt
     3721 gcccgtgctg atcgagctgg acggcgacgt gcacggccac aagttcagcg tgcgcggcga
     3781 gggcgagggc gacgccgact acggcaagct ggagatcaag ttcatctgca ccaccggcaa
     3841 gctgcccgtg ccctggccca ccctggtgac caccctctgc tacggcatcc agtgcttcgc
     3901 ccgctacccc gagcacatga agatgaacga cttcttcaag agcgccatgc ccgagggcta
     3961 catccaggag cgcaccatcc agttccagga cgacggcaag tacaagaccc gcggcgaggt
     4021 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcaagg acttcaagga
     4081 ggacggcaac atcctgggcc acaagctgga gtacagcttc aacagccaca acgtgtacat
     4141 ccgccccgac aaggccaaca acggcctgga ggctaacttc aagacccgcc acaacatcga
     4201 gggcggcggc gtgcagctgg ccgaccacta ccagaccaac gtgcccctgg gcgacggccc
     4261 cgtgctgatc cccatcaacc actacctgag cactcagacc aagatcagca aggaccgcaa
     4321 cgaggcccgc gaccacatgg tgctcctgga gtccttcagc gcctgctgcc acacccacgg
     4381 catggacgag ctgtacaggt ccggactcag ataagtttaa acggccggcc gcggtctgta
     4441 caagtaggat tcgtcgaggg acctaataac ttcgtatagc atacattata cgaagttata
     4501 catgtttaag ggttccggtt ccactaggta caattcgata tcaagcttat cgataatcaa
     4561 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4621 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4681 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4741 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4801 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4861 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4921 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4981 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5041 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     5101 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     5161 cgatcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     5221 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     5281 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     5341 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     5401 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     5461 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     5521 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     5581 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     5641 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     5701 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     5761 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     5821 acccttttag tcagtgtgga aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc
     5881 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5941 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6001 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6061 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6121 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6181 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6241 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6301 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6361 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6421 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6481 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6541 gtggaacgaa aactcacgtt aagggatttt ggtcatgatt acgccccgcc ctgccactca
     6601 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     6661 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     6721 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     6781 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     6841 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     6901 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     6961 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     7021 aactccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     7081 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     7141 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     7201 atatcaacgg tggtatatcc agtgattttt ttctccatac tcttcctttt tcaatattat
     7261 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7321 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
