LOCUS       sequence-203305-        6507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gene expression of FLAGHA-TIA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6507)
  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 6507)
  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_FlagHA_TIA1
FEATURES             Location/Qualifiers
     source          1..6507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(60..648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(140..159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(819..1610,1611..1679))
                     /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     1442..1461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1680..1784)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1859..1878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2050..2429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2430..2633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2584..2604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2635..2653
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2656..2674
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2684..2708
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2808..2831
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2832..2858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2874..2898
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4076..4100)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4153..4194
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(4274..4291)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4280..4504
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4637..4656)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4788..4835
                     /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             4843..5865
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4843..5865
                     /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    5995..6116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6032..6051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6086..6105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6165..6181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6165..6181)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6178..6200)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6189..6205
                     /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(6213..6219,6220..6237,6238..6243))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6258..6279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6396..6413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
       61 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      481 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
      841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
      961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     1141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     1441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     1801 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     1861 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     1921 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     1981 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     2041 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     2101 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2161 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2221 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2281 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2341 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2401 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2461 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2521 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2581 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctcccta
     2641 tcagtgatag agatctccct atcagtgata gagatcgtcg acgagctcgt ttagtgaacc
     2701 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
     2761 gatccagcct ccggactcta gcgtttaaac ttaagcttgg taccatggac tacaaggacg
     2821 acgatgacaa gtacccttat gacgtgcccg attacgcttt gaaccagata tcaacaagtt
     2881 tgtacaaaaa agcaggctcc accatgggaa ccaattcagt cgacatggag gacgagatgc
     2941 ccaagactct atacgtcggt aacctttcca gagatgtgac agaagctcta attctgcaac
     3001 tctttagcca gattggacct tgtaaaaact gcaaaatgat tatggataca gctggaaatg
     3061 atccctattg ttttgtggag tttcatgagc atcgtcatgc agctgcagca ttagctgcta
     3121 tgaatggacg gaagataatg ggtaaggaag tcaaagtgaa ttgggcaaca acccctagca
     3181 gtcaaaagaa agatacaagc aatcatttcc atgtctttgt tggtgatctc agcccagaaa
     3241 ttacaactga agatataaaa gctgcttttg caccatttgg aagaatatca gatgcccgag
     3301 tggtaaaaga catggcaaca ggaaagtcta agggatatgg ctttgtctcc tttttcaaca
     3361 aatgggatgc tgaaaacgcc attcaacaga tgggtggcca gtggcttggt ggaagacaaa
     3421 tcagaactaa ctgggcaacc cgaaagcctc ccgctccaaa gagtacatat gagtcaaata
     3481 ccaaacagct atcatatgat gaggttgtaa atcagtctag tccaagcaac tgtactgtat
     3541 actgtggagg tgttacttct gggctaacag aacaactaat gcgtcagact ttttcaccat
     3601 ttggacaaat aatggaaatt cgagtctttc cagataaagg atattcattt gttcggttca
     3661 attcccatga aagtgcagca catgcaattg tttctgttaa tggtactacc attgaaggtc
     3721 atgttgtgaa atgctattgg ggcaaagaaa ctcttgatat gataaatccc gtgcaacagc
     3781 agaatcaaat tggatatccc caaccttatg gccagtgggg ccagtggtat ggaaatgcac
     3841 aacaaattgg ccagtatatg cctaatggtt ggcaagttcc tgcatatgga atgtatggcc
     3901 aggcatggaa ccagcaagga tttaatcaga cacagtcttc tgcaccatgg atgggaccaa
     3961 attatggagt gcaaccgcct caagggcaaa atggcagcat gttgcccaat cagccttctg
     4021 ggtatcgagt ggcagggtat gaaacccagt gagcggccgc actcgagata tctagaccca
     4081 gctttcttgt acaaagtggt tgatatccag cacagtggcg gccgctcaag tctagagggc
     4141 ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc
     4201 tgatccgcgg ccgcatagat aactgatcca gtgtgcctcg agtctagagg gcccgtttaa
     4261 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4321 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4381 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4441 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4501 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4561 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4621 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4681 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4741 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa gttcctattc
     4801 cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc tgaactcacc
     4861 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     4921 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     4981 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     5041 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg
     5101 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     5161 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
     5221 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     5281 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     5341 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     5401 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     5461 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     5521 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     5581 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
     5641 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     5701 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     5761 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     5821 agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt actacgagat
     5881 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5941 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6001 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6061 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6121 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6181 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6241 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6301 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6361 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6421 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6481 gaatcagggg ataacgcagg aaagaac
//
