LOCUS       sequence-244101-        8607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ATF4 expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8607)
  AUTHORS   Bagheri-Yarmand R, Sinha KM, Gururaj AE, Ahmed Z, Rizvi YQ, Huang 
            SC, Ladbury JE, Bogler O, Williams MD, Cote GJ, Gagel RF
  TITLE     A novel dual kinase function of the RET proto-oncogene negatively 
            regulates activating transcription factor 4-mediated apoptosis.
  JOURNAL   J Biol Chem. 2015 May 1;290(18):11749-61. doi: 
            10.1074/jbc.M114.619833. Epub 2015 Mar 20.
  PUBMED    25795775
REFERENCE   2  (bases 1 to 8607)
  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: 
            "10.1074/jbc.M114.619833"; journalName: "J Biol Chem"; date: 
            "2015-05-1- 1"; volume: "290"; issue: "18"; pages: "11749-61"
COMMENT     Sequence Label: plenti-ATF4
FEATURES             Location/Qualifiers
     source          1..8607
                     /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"
     regulatory      3285..3294
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3294..3317
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4130..4147
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="GSSKSK"
     CDS             4393..4422
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4441..4464
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4521..5073
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4665..4682)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4892..4911
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5073..5672
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5073..5092)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5569..5589
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(5702..5735)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5791..6379
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5844..5864)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6262..6273)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6382..6398)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6383..6399)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6908..7088
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7150..7738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7230..7249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7815..8474)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     8389..8408
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(8475..8579)
                     /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 attcgccgcc atggactaca
     3301 aggacgacga tgacaagggg tcgaccatga ccgaaatgag cttcctgagc agcgaggtgt
     3361 tggtggggga cttgatgtcc cccttcgacc agtcgggttt gggggctgaa gaaagcctag
     3421 gtctcttaga tgattacctg gaggtggcca agcacttcaa acctcatggg ttctccagcg
     3481 acaaggctaa ggcgggctcc tccgaatggc tggctgtgga tgggttggtc agtccctcca
     3541 acaacagcaa ggaggatgcc ttctccggga cagattggat gttggagaaa atggatttga
     3601 aggagttcga cttggatgcc ctgttgggta tagatgacct ggaaaccatg ccagatgacc
     3661 ttctgaccac gttggatgac acttgtgatc tctttgcccc cctagtccag gagactaata
     3721 agcagccccc ccagacggtg aacccaattg gccatctccc agaaagttta acaaaacccg
     3781 accaggttgc ccccttcacc ttcttacaac ctcttcccct ttccccaggg gtcctgtcct
     3841 ccactccaga tcattccttt agtttagagc tgggcagtga agtggatatc actgaaggag
     3901 ataggaagcc agactacact gcttacgttg ccatgatccc tcagtgcata aaggaggaag
     3961 acaccccttc agataatgat agtggcatct gtatgagccc agagtcctat ctggggtctc
     4021 ctcagcacag cccctctacc aggggctctc caaataggag cctcccatct ccaggtgttc
     4081 tctgtgggtc tgcccgtccc aaaccttacg atcctcctgg agagaagatg gtagcagcaa
     4141 aagtaaaggg tgagaaactg gataagaagc tgaaaaaaat ggagcaaaac aagacagcag
     4201 ccactaggta ccgccagaag aagagggcgg agcaggaggc tcttactggt gagtgcaaag
     4261 agctggaaaa gaagaacgag gctctaaaag agagggcgga ttccctggcc aaggagatcc
     4321 agtacctgaa agatttgata gaagaggtcc gcaaggcaag ggggaagaaa agggtcccct
     4381 aggcggccgc tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg
     4441 gattacaagg atgacgacga taaggtttaa acctaggcgt agcggccgca aattccgccc
     4501 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     4561 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     4621 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     4681 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     4741 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     4801 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     4861 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     4921 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     4981 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     5041 cgtggttttc ctttgaaaaa cacgatgatc atatgaccga gtacaagccc acggtgcgcc
     5101 tcgccacccg cgacgacgtc ccccgggcag tacgcaccct cgccgccgcg ttcgccgact
     5161 accccgccac gcgccacacc gtcgatccag accgccacat cgagcgggtc accgagctgc
     5221 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5281 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5341 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5401 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5461 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5521 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5581 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5641 cctggtgcat gacccgcaag cccggtgcct gatgtacaag taggattcgt cgagggacct
     5701 aataacttcg tatagcatac attatacgaa gttatacatg tttaagggtt ccggttccac
     5761 taggtacaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     5821 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5881 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5941 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     6001 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     6061 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6121 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6181 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6241 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6301 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6361 ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg
     6421 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
     6481 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
     6541 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     6601 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt
     6661 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg
     6721 gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa
     6781 cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt
     6841 agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga
     6901 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6961 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     7021 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     7081 ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7561 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7681 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7741 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7801 gattttggtc atgattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     7861 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     7921 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     7981 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     8041 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     8101 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     8161 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     8221 catatcacca gctcaccgtc tttcattgcc atacggaact ccggatgagc attcatcagg
     8281 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     8341 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     8401 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     8461 atttttttct ccatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8521 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8581 acatttcccc gaaaagtgcc acctgac
//
