LOCUS       sequence-289552-        7975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Fluc-RRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7975)
  AUTHORS   Teplova M, Wohlbold L, Khin NW, Izaurralde E, Patel DJ
  TITLE     Structure-function studies of nucleocytoplasmic transport of 
            retroviral genomic RNA by mRNA export factor TAP.
  JOURNAL   Nat Struct Mol Biol. 2011 Aug 7;18(9):990-8. doi: 10.1038/nsmb.2094.
  PUBMED    21822283
REFERENCE   2  (bases 1 to 7975)
  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/nsmb"; 
            journalName: "Nat Struct Mol Biol"; date: "2011-08-7- 7"; volume: 
            "18"; issue: "9"; pages: "990-8"
COMMENT     Sequence Label: pCMV128-Fluc-RRE_O
FEATURES             Location/Qualifiers
     source          1..7975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        2338..2717
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2719..2918
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2872..2892
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3301..3310
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3307..4959
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3373..3391)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    6142..6375
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6560..6604
                     /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"
     primer_bind     complement(7581..7598)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7581..7597)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7590..7612)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7806..7825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7925..7947
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcctgc aggtcgaggt cgacccattg catacgttgt
     2281 atccatatca taatatgtac atttatattg gctcatgtcc aacattaccg ccatgttgac
     2341 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2401 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2461 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2521 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2581 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2641 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2701 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2761 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2821 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2881 gcggtaggcg tgtacggtgg gaggtctata taagcagagt ctagagccct ggaagcatcc
     2941 aggaagtcag cctaggactg cttgtaacaa ttgctattgt aaaaagtgtt gctttcattg
     3001 ctacgcgtgt ttcacaagaa aaggcttagg catctccttg atcaggaaga agcggagaca
     3061 gcgacgaaga gctcctcagg acagtcagac tcatcaagct tctctatcaa agcagtaagt
     3121 agtaaatgta atgcaatctt tacaaatatt agcaatagta tcattagtag tagtagcaat
     3181 aatagcaata gttgtgtgga ccatagtact catagaatat aggaaaatat taagacaaag
     3241 aaaatagaca gattaattga tagaataaga gaaaaagcag aagacagtgg cggccgcaca
     3301 gccaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa
     3361 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga
     3421 acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa
     3481 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc
     3541 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc
     3601 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg
     3661 ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac
     3721 gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa aacggattac
     3781 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa
     3841 tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc
     3901 tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga
     3961 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta
     4021 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt
     4081 ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag
     4141 gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc
     4201 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc
     4261 ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa
     4321 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa
     4381 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc
     4441 gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg
     4501 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta
     4561 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg
     4621 aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg
     4681 ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt
     4741 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc
     4801 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt
     4861 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc
     4921 ctcataaagg ccaagaaggg cggaaagatc gccgtgtaag gatcctctag tctgtgttac
     4981 tttaaattgc actgatttgg ggaaggctac taataccaat agtagtaatt ggaaagaaga
     5041 aataaaagga gaaataaaaa actgctcttt caatatcacc acaagcataa gagataagat
     5101 tcagaaagaa aatgcacttt ttcgtaacct tgatgtagta ccaatagata atgctagtac
     5161 tactaccaac tataccaact ataggttgat acattgtaac agatcagtca ttacacaggc
     5221 ctgtccaaag gtatcatttg agccaattcc catacattat tgtaccccgg ctggttttgc
     5281 gattctaaag tgtaataata aaacgttcaa tggaaaagga ccatgtacaa atgtcagcac
     5341 agtacaatgt acacatggaa ttaggccaat agtgtcaact caactgctgt taaatggcag
     5401 tctagcagaa gaagaggtag taattagatc tgacaatttc acgaacaatg ctaaaaccat
     5461 aatagtacag ctgaatgaat ctgtagcaat taactgtaca agacccaaca acaatacaag
     5521 aaaaagtatc tatataggac cagggagagc atttcataca acaggaagaa taataggaga
     5581 tataagaaaa gcacattgta acattagtag agcacaatgg aataacactt tagaacagat
     5641 agttaaaaaa ttaagagaac agtttgggaa taataaaaca atagtcttta atcaatcctc
     5701 aggaggggac ccagaaattg taatgcacag ttttaattgt agaggggaat ttttctactg
     5761 taatacaaca caactgttta ataatacatg gaggttaaat cacactgaag gaactaaagg
     5821 aaatgacaca atcatactcc catgtagaat aaaacaaatt ataaacatgt ggcaggaagt
     5881 aggaaaagca atgtatgccc ctcccattgg aggacaaatt agttgttcat caaatattac
     5941 agggctgcta ttaacaagag atggtggtac aaatgtaact aatgacaccg aggtcttcag
     6001 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtaat
     6061 aaaaattgaa ccattaggaa tagcacccac caaggcaaag agaagagtgg tgcagagaga
     6121 aaaaagagca gtgggaatag taggagctat gttccttggg ttcttgggag cagcaggaag
     6181 cactatgggc gcagtgtcat tgacgctgac ggtacaggcc agacaattat tgtctggtat
     6241 agtgcaacag cagaacaatt tgctgagggc tattgaggcg caacaacatc tgttgcaact
     6301 cacagtctgg ggcatcaagc agctccaggc aagagtcctg gctgtggaaa gatacctaag
     6361 ggatcaacag ctcctaggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     6421 gccttggaat gctagttgga gtaataaatc tctggaagac atttgggata acatgacctg
     6481 gatgcagtgg gaaagagaaa ttgacaatta cacaaacaca atatacacct tacttgaaga
     6541 atcgcagaac caacaagaaa agaatgaaca agaattatta gaattggata agtgggcaag
     6601 tttgtggaat tggtttagca taacaaactg gctgtggtat ataaagatat tcataatgat
     6661 agtaggaggc ttggtaggtt taagaatagt ttttgctgtg ctttctatag tgaatagagt
     6721 taggcaggga tactcaccat tgtcatttca gacccgcctc ccagtcctcg agacctggaa
     6781 aaacatggag caatcacaag tagcaataca gcagctacta atgctgattg tgcctggcta
     6841 gaagcacaag aggaggaaga ggtgggtttt ccagtcagac ctcaggtacc tttaagacca
     6901 atgacttaca aggcagcttt agatattagc cactttttaa aagaaaaggg gggactggaa
     6961 gggctaattt ggtcccaaag aagacaagag atccttgatc tgtggatcta ccacacacaa
     7021 ggctacttcc ctgattggca gaattacaca ccagggccag ggatcagata tccactgacc
     7081 tttggatggt gcttcaagct agtaccagtt gagccagaga aggtagaaga ggccaatgaa
     7141 ggagagaaca acagcttgtt acaccctatg agcctgcatg ggatggagga cgcggagaaa
     7201 gaagtgttag tgtggaggtt tgacagcaaa ctagcatttc atcacatggc ccgagagctg
     7261 catccggagt actacaaaga ctgctgacat cgagctttct acaagggact ttccgctggg
     7321 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgtccctc agatgctgca
     7381 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     7441 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7501 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7561 ttttagtcag tgtggaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     7621 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     7681 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     7741 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     7801 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     7861 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     7921 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
