LOCUS       sequence-203571-        7405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression of EGFP - CELF1
             .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7405)
  AUTHORS   Meyer C, Garzia A, Mazzola M, Gerstberger S, Molina H, Tuschl T
  TITLE     The TIA1 RNA-Binding Protein Family Regulates EIF2AK2-Mediated 
            Stress Response and Cell Cycle Progression.
  JOURNAL   Mol Cell. 2018 Feb 15;69(4):622-635.e6. doi: 
            10.1016/j.molcel.2018.01.011.
  PUBMED    29429924
REFERENCE   2  (bases 1 to 7405)
  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: 
            "10.1016/j.molcel.2018.01"; journalName: "Mol Cell"; date: 
            "2018-02-15- 15"; volume: "69"; issue: "4"; pages: "622-635.e6"
COMMENT     Sequence Label: pFRT_TO_eGFP_CELF1
FEATURES             Location/Qualifiers
     source          1..7405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(756..758,759..761,762..1472)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(756..758,759..761,762..1472)
                     /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(801..822)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1062..1081)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1409..1430
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1479..1503
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2967..2991)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3044..3085
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3123..3140)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3129..3353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3486..3505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3637..3684
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3692..4714
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3692..4714
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4844..4965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4881..4900)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4935..4954
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5014..5030)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5014..5030)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5027..5049)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5038..5054
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5062..5068,5069..5086,5087..5092))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5107..5128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5245..5262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5416..6004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5496..5515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6175..6966,6967..7035))
                     /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     6798..6817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7036..7140)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7215..7234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccatggt gagcaagggc gaggagctgt
      781 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      841 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      901 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      961 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1021 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1081 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1141 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1201 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     1261 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1321 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     1381 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1441 ggatcactct cggcatggac gagctgtaca agaagcttac aagtttgtac aaaaaagcag
     1501 gcttcatgaa cggcaccctg gaccacccag accaaccaga tcttgatgct atcaagatgt
     1561 ttgtgggcca ggttccaagg acctggtctg aaaaggactt gcgggaactc ttcgaacagt
     1621 atggtgctgt gtatgaaatc aacgtcctaa gggataggag ccaaaacccg cctcagagca
     1681 aagggtgctg ttttgttaca ttttacaccc gtaaagctgc attagaagct cagaatgctc
     1741 ttcacaacat gaaagtcctc ccagggatgc atcaccctat acagatgaaa cctgctgaca
     1801 gtgagaagaa caatgcagtg gaagacagga agctgtttat tggtatgatt tccaagaagt
     1861 gcactgaaaa tgacatccga gtcatgttct cttcgtttgg acagattgaa gaatgccgga
     1921 tattgcgggg acctgatggc ctgagccgag gttgtgcatt tgtgactttt acaacaagag
     1981 ccatggcaca gacggctatc aaggcaatgc accaagcaca gaccatggag ggttgctcat
     2041 cacccatggt ggtaaaattt gctgatacac agaaggacaa agaacagaag agaatggccc
     2101 agcagctcca gcagcagatg cagcaaatca gcgcagcatc tgtgtgggga aaccttgctg
     2161 gtctaaatac tcttggaccc cagtatttag cactttattt gcagctcctt cagcagactg
     2221 cctcctctgg gaacctcaac accctgagca gcctccaccc aatgggaggg ttgaatgcaa
     2281 tgcagttaca gaatttggct gcactagctg ctgcagctag tgcagctcag aacacaccaa
     2341 gtggtaccaa tgctctcact acatccagca gtcccctcag cgtgctcact agttcagggt
     2401 cctcacctag ctctagcagc agtaattctg tcaaccccat agcctcactt ggagccctgc
     2461 agacattagc tggagcaacg gctggcctca atgttggctc tttggcagga atggctgctt
     2521 taaatggtgg cctgggcagc agtggccttt ccaatggcac cgggagcacc atggaggccc
     2581 tcactcaggc ctactcgggt atccagcaat atgctgctgc tgcgctcccc actctgtaca
     2641 accagaatct tctgacacag cagagtattg gtgctgctgg aagccagaag gaaggtccag
     2701 agggagccaa cctgttcatc taccacctgc cccaggagtt tggtgatcag gacctgctgc
     2761 agatgtttat gccctttggg aatgtcgtgt ctgccaaggt tttcatagac aagcagacaa
     2821 acctgagcaa gtgttttggt tttgtaagtt acgacaatcc tgtttcggcc caagctgcca
     2881 tccagtccat gaacggcttt cagattggca tgaagcggct taaagtgcag ctcaaacgtt
     2941 cgaagaatga cagcaagccc tactgaaccc agctttcttg tacaaagtgg ttgatatcca
     3001 gcacagtggc ggccgctcaa gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
     3061 ccctctcctc ggtctcgatt ctacgctcga gtctagaggg cccgtttaaa cccgctgatc
     3121 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3181 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3241 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3301 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3361 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3421 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3481 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3541 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3601 caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc gaagttccta
     3661 ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg cgacgtctgt
     3721 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     3781 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     3841 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     3901 gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat
     3961 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     4021 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag
     4081 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4141 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4201 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4261 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4321 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4381 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4441 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
     4501 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4561 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4621 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4681 acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac
     4741 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4801 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4861 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4921 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4981 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5041 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5101 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5161 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5221 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5281 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5341 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5821 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6241 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7141 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7201 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7261 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7321 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     7381 gatgtacggg ccagatatac gcgtt
//
