LOCUS       Exported                7153 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses hFXR1.
ACCESSION   .
VERSION     .
KEYWORDS    pFRT-TODestFLAGHAhFXR1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  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 7153)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7153
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        182..561
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        562..765
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     716..736
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    767..785
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    788..806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     816..840
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             940..963
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             964..990
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1006..1030
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2854..2878)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2931..2972
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3052..3069)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3058..3282
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3415..3434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3566..3613
                     /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             3621..4643
                     /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    4773..4894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4810..4829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4864..4883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4943..4959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4943..4959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4956..4978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4967..4983
                     /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(4991..5021)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5036..5057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5174..5191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5345..5933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5425..5444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6104..6964)
                     /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     6727..6746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6965..7069)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7144..10)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
       61 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      121 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
      181 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      241 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      301 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      361 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      421 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      481 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      541 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      601 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      661 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      721 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctccc tatcagtgat
      781 agagatctcc ctatcagtga tagagatcgt cgacgagctc gtttagtgaa ccgtcagatc
      841 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
      901 ctccggactc tagcgtttaa acttaagctt ggtaccatgg actacaagga cgacgatgac
      961 aagtaccctt atgacgtgcc cgattacgct ttgaaccaga tatcaacaag tttgtacaaa
     1021 aaagcaggct ccaccatggg aaccaattca gtcgacatgg cggagctgac ggtggaggtt
     1081 cgcggctcta acggggcttt ctacaaggga tttatcaaag atgttcatga agactccctt
     1141 acagttgttt ttgaaaataa ttggcaacca gaacgccagg ttccatttaa tgaagttaga
     1201 ttaccaccac cacctgatat aaaaaaagaa attagtgaag gagatgaagt agaggtatat
     1261 tcaagagcaa atgaccaaga gccatgtggg tggtggttgg ctaaagttcg gatgatgaaa
     1321 ggagaatttt atgtcattga atatgctgct tgtgacgcta cttacaatga aatagtcaca
     1381 tttgaacgac ttcggcctgt caatcaaaat aaaactgtca aaaaaaatac cttctttaaa
     1441 tgcacagtgg atgttcctga ggatttgaga gaggcgtgtg ctaatgaaaa tgcacataaa
     1501 gattttaaga aagcagtagg agcatgcaga attttttacc atccagaaac aacacagcta
     1561 atgatactgt ctgccagtga agcaactgtg aagagagtaa acatcttaag tgacatgcat
     1621 ttgcgaagta ttcgtacgaa gttgatgctt atgtccagaa atgaagaggc cactaagcat
     1681 ttagaatgca caaaacaact tgcagcagct tttcatgagg aatttgttgt gagagaagat
     1741 ttaatgggcc tggcaatagg aacacatggt agtaacatcc agcaagctag gaaggttcct
     1801 ggagttaccg ccattgagct agatgaagat actggaacat tcagaatcta cggagagagt
     1861 gctgatgctg taaaaaaggc tagaggtttc ttggaatttg tggaggattt tattcaggtt
     1921 cctaggaatc tcgttggaaa agtaattgga aaaaatggca aagttattca agaaatagtg
     1981 gacaaatctg gtgtggttcg agtgagaatt gaaggggaca atgaaaataa attacccaga
     2041 gaagacggta tggttccatt tgtatttgtt ggcactaaag aaagcattgg aaatgtgcag
     2101 gttcttctag agtatcatat tgcctatcta aaggaagtag aacagctaag aatggaacgc
     2161 ctacagattg atgaacagct gcgacagatt ggttctaggt cttatagcgg aagaggcaga
     2221 ggtcgtcggg gacctaatta cacctccggt tatggtacaa attctgagct gtctaacccc
     2281 tctgaaacgg aatctgagcg taaagacgag ctgagtgatt ggtcattggc aggagaagat
     2341 gatcgagaca gccgacatca gcgtgacagc aggagacgcc caggaggaag aggcagaagt
     2401 gtttcagggg gtcgaggtcg tggtggacca cgtggtggca aatcctccat cagttctgtg
     2461 ctcaaagatc cagacagcaa tccatacagc ttacttgata atacagaatc agatcagact
     2521 gcagacactg atgccagcga atctcatcac agtactaacc gtcgtaggcg gtctcgtaga
     2581 cgaaggactg atgaagatgc tgttctgatg gatggaatga ctgaatctga tacagcttca
     2641 gttaatgaaa atgggctagg caaaagatgt gattgaagag catggtcctt cagaaaaggc
     2701 aataaacggc ccaactagtg cttctggcga tgacatttct aagctacagc gtactccagg
     2761 agaagaaaag attaatacct taaaagaaga aaacactcaa gaagcagcag tcctgaatgg
     2821 tgtttcataa gcggccgcac tcgagatatc tagacccagc tttcttgtac aaagtggttg
     2881 atatccagca cagtggcggc cgctcaagtc tagagggccc gcggttcgaa ggtaagccta
     2941 tccctaaccc tctcctcggt ctcgattcta cgcgtacctg atccgcggcc gcatagataa
     3001 ctgatccagt gtgcctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     3061 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3121 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3181 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3241 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3301 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3361 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3421 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3481 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3541 attagggtga tggttcacgt acctagaagt tcctattccg aagttcctat tctctagaaa
     3601 gtataggaac ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     3661 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     3721 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     3781 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     3841 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     3901 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     3961 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     4021 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     4081 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     4141 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     4201 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     4261 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     4321 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     4381 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     4441 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     4501 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     4561 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     4621 ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct
     4681 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4741 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4801 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4861 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4921 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4981 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5041 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5101 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5761 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5881 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6001 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6061 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6121 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6181 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6241 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6301 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6361 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6421 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6481 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6541 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6601 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6661 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6721 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6781 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6841 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6901 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6961 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7021 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7081 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7141 cagtacaatc tgc
//
