LOCUS       sequence-83256-e        7443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hFXR2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7443)
  AUTHORS   Ascano M Jr, Mukherjee N, Bandaru P, Miller JB, Nusbaum JD, Corcoran
            DL, Langlois C, Munschauer M, Dewell S, Hafner M, Williams Z, Ohler 
            U, Tuschl T
  TITLE     FMRP targets distinct mRNA sequence elements to regulate protein 
            expression.
  JOURNAL   Nature. 2012 Dec 20;492(7429):382-6. doi: 10.1038/nature11737. Epub 
            2012 Dec 12.
  PUBMED    23235829
REFERENCE   2  (bases 1 to 7443)
  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/nature11737"; journalName: "Nature"; date: "2012-12-20- 
            20"; volume: "492"; issue: "7429"; pages: "382-6"
COMMENT     Sequence Label: pFRT-TODestFLAGHAhFXR2
FEATURES             Location/Qualifiers
     source          1..7443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..42
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(86..105)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        277..656
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        657..860
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     811..831
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    862..880
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    883..901
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     911..935
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1035..1058
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1059..1085
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1101..1125
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3197..3221)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3274..3315
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3395..3412)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3401..3625
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3758..3777)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3909..3956
                     /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             3964..4986
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3964..4986
                     /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    5116..5237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5153..5172)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5207..5226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5286..5302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5286..5302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5299..5321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5310..5326
                     /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(5334..5340,5341..5358,5359..5364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5379..5400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5517..5534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5688..6276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5768..5787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6447..7238,7239..7307))
                     /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     7070..7089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7308..7412)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtaagccta tccctaaccc tctcctcggt ctcgattcta cggacggatc gggagatctc
       61 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
      121 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
      181 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
      241 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
      301 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      361 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      421 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      481 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      541 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      601 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      661 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      721 gtctccaccc cattgacgtc aatgggagtt tgttttggaa ccaaaatcaa cgggactttc
      781 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      841 aggtctatat aagcagagct ctccctatca gtgatagaga tctccctatc agtgatagag
      901 atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
      961 ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcg tttaaactta
     1021 agcttggtac catggactac aaggacgacg atgacaagta cccttatgac gtgcccgatt
     1081 acgctttgaa ccagatatca acaagtttgt acaaaaaagc aggctccacc atgggaacca
     1141 attcagtcga catgggcggc ctggcctctg ggggggatgt ggagccggga ctgcccgtcg
     1201 aggtgcgcgg ctccaacggg gccttctaca agggctttgt gaaggatgtc catgaagact
     1261 ctgtcaccat cttctttgaa aacaactggc agagtgagag acaaattcct tttggggatg
     1321 tccggctacc acctccagct gactataata aggagatcac agaaggggat gaagtggagg
     1381 tttattctcg agccaatgaa caggaacctt gtggctggtg gctggcccgg gtgcggatga
     1441 tgaagggaga tttctatgtc attgaatatg ctgcctgtga tgccacctac aatgaaattg
     1501 ttaccctgga gcgacttcgg ccagttaatc ccaatcccct tgcaaccaaa ggcagcttct
     1561 tcaaggttac catggctgtg cccgaggatc tgagagaagc ctgctccaat gaaaacgtcc
     1621 ataaagagtt caagaaagcc ctgggagcca actgcatctt tctcaacatc acaaacagtg
     1681 agctcttcat tctgtcaacc acagaagccc ctgtgaagcg agcatctctg ctgggtgata
     1741 tgcatttccg aagcctgcgc accaaactgc tacttatgtc ccgcaatgaa gaagctacca
     1801 agcacctaga gacaagcaag cagttggcag cagccttcca agaggagttc acagtgcgag
     1861 aggacctgat gggactggca attgggactc acggtgccaa catccagcag gcccgaaaag
     1921 tacctggggt gaccgccatt gagttgggtg aagagacctg cactttccgc atctatgggg
     1981 agactcccga ggcttgccga caggcccgaa gctaccttga gttttctgag gactcagtgc
     2041 aagtgcccag gaacctggtt ggcaaagtga ttggaaagaa cgggaaagtg atccaggaga
     2101 ttgtggataa atctggtgtg gtgagggttc gagtggaagg tgataatgac aagaagaacc
     2161 ccagggagga gggaatggtt cccttcattt ttgttggcac ccgagagaac atcagcaatg
     2221 cccaggcttt gctggagtat cacctctcct acctgcagga ggtagagcag cttcgcttgg
     2281 agaggctaca aattgatgag cagcttcggc agattgggct gggctttcgc cctcctggga
     2341 gtgggcgggg cagcggtggc agcgacaagg ctggatatag cactgatgag agctcctcct
     2401 cctccctcca tgcgactcga acctatgggg gcagctatgg gggccgtggc cgtggccgga
     2461 ggacaggcgg tcctgcctat ggccccagct cagatgtgtc tacagcttca gagactgagt
     2521 cagagaagag agaggagccc aaccgagctg ggcctggcga cagggatccc ccaacccgag
     2581 gggaagaaag ccggaggcgg ccgactgggg gccggggtag gggaccccca cctgcccccc
     2641 ggcccacttc gagatacaat tcttcatcta ttagctcagt gctgaaggat ccagacagta
     2701 atccctacag cctattggac acgtctgaac cagagccccc ggttgattca gaacctgggg
     2761 aacccccccc agcaagtgcc aggcgccgcc gctcccgccg ccgccgcact gatgaagaca
     2821 ggaccgtcat ggatggaggc ctggaatcag atgggcccaa catgacagag aatggcctgg
     2881 aagatgaatc aagacctcaa cgtcgtaatc gcagccgccg ccgccgtaac cgtggtaatc
     2941 ggactgatgg ctctatcagt ggagaccgcc agccagtgac tgtggctgac tatatctcac
     3001 gagcagagtc tcagagccgc cagaggccac ccctggaacg cactaaaccc tcagaagact
     3061 ctctttcagg acagaagggt gactctgtca gcaagcttcc taagggcccc tcggagaatg
     3121 gggagctctc cgcccccttg gagttgggta gtatggtgaa tggggtttca taagcggccg
     3181 cactcgagat atctagaccc agctttcttg tacaaagtgg ttgatatcca gcacagtggc
     3241 ggccgctcaa gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc
     3301 ggtctcgatt ctacgcgtac cggatccgcg gccgcataga taactgatcc agtgtgcctc
     3361 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     3421 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3481 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3541 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3601 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3661 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3721 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3781 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3841 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3901 cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg aacttccttg
     3961 gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     4021 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     4081 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     4141 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     4201 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     4261 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     4321 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     4381 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     4441 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     4501 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     4561 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     4621 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     4681 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     4741 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     4801 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     4861 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     4921 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     4981 gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5041 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5101 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5161 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5221 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5281 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5401 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5461 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5521 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5581 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5641 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5701 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5761 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5821 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5881 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5941 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6001 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6061 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6121 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6421 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6481 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6541 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6601 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6661 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6721 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6781 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6841 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6901 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6961 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7021 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7081 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7141 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7201 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7261 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7321 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7381 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7441 gtc
//
