LOCUS       sequence-409993-        7263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Rluc-RRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7263)
  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 7263)
  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-Rluc-RRE_O
FEATURES             Location/Qualifiers
     source          1..7263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..474
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        475..678
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     629..649
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1067..1076
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1073..2008
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    3188..3421
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3606..3650
                     /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(4629..4646)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4629..4645)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4638..4660)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4854..4873)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4973..4995
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5033..5051)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5119..5223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5224..5292,5293..6084)
                     /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(5442..5461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6255..6843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6744..6763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6997..7014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7131..7152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7167..7172,7173..7190,7191..7197)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7205..7221
                     /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     7210..7232
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7229..7245
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7229..7245
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcatgcctgc aggtcgaaat tccattgcat acgttgtatc tatatcataa tatgtacatt
       61 tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa
      121 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      181 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      241 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      301 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
      361 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      421 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      481 cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      541 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      601 aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag
      661 gtctatataa gcagagctcg gtacccctag agccctggaa gcatccagga agtcagccta
      721 ggactgcttg taacaattgc tattgtaaaa agtgttgctt tcattgctac gcgtgtttca
      781 caagaaaagg cttaggcatc tccttgatca ggaagaagcg gagacagcga cgaagagctc
      841 ctcaggacag tcagactcat caagcttctc tatcaaagca gtaagtagta aatgtaatgc
      901 aatctttaca aatattagca atagtatcat tagtagtagt agcaataata gcaatagttg
      961 tgtggaccat agtactcata gaatatagga aaatattaag acaaagaaaa tagacagatt
     1021 aattgataga ataagagaaa gagcagaaga cagtggcggc cgcacagcca ccatggcttc
     1081 caaggtgtac gaccccgagc aacgcaaacg catgatcact gggcctcagt ggtgggctcg
     1141 ctgcaagcaa atgaacgtgc tggactcctt catcaactac tatgattccg agaagcacgc
     1201 cgagaacgcc gtgatttttc tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt
     1261 cgtgcctcac atcgagcccg tggctagatg catcatccct gatctgatcg gaatgggtaa
     1321 gtccggcaag agcgggaatg gctcatatcg cctcctggat cactacaagt acctcaccgc
     1381 ttggttcgag ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg actggggggc
     1441 ttgtctggcc tttcactact cctacgagca ccaagacaag atcaaggcca tcgtccatgc
     1501 tgagagtgtc gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg aggaggatat
     1561 cgccctgatc aagagcgaag agggcgagaa aatggtgctt gagaataact tcttcgtcga
     1621 gaccatgctc ccaagcaaga tcatgcggaa actggagcct gaggagttcg ctgcctacct
     1681 ggagccattc aaggagaagg gcgaggttag acggcctacc ctctcctggc ctcgcgagat
     1741 ccctctcgtt aagggaggca agcccgacgt cgtccagatt gtccgcaact acaacgccta
     1801 ccttcgggcc agcgacgatc tgcctaagat gttcatcgag tccgaccctg ggttcttttc
     1861 caacgctatt gtcgagggag ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg
     1921 cctccacttc agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt
     1981 ggagcgcgtg ctgaagaacg agcagtaagg atcctctagg tgttacttta aattgcactg
     2041 atttggggaa ggctactaat accaatagta gtaattggaa agaagaaata aaaggagaaa
     2101 taaaaaactg ctcttttaat atcaccacaa gcataagaga taagattcag aaagaaaatg
     2161 cactttttcg taaccttgat gtagtaccaa tagataatgc tagtactact accaactata
     2221 ccaactatag gttgatacat tgtaacagat cagtcattac acaggcctgt ccaaaggtat
     2281 catttgagcc aattcccata cattattgta ccccggctgg ttttgcgatt ctaaagtgta
     2341 ataataaaac gttcaatgga aaaggaccat gtacaaatgt cagcacagta caatgtacac
     2401 atggaattag gccaatagtg tcaactcaac tgctgttaaa tggcagtcta gcagaagaag
     2461 aggtagtaat tagatctgac aatttcacga acaatgctaa aaccataata gtacagctga
     2521 atgaatctgt agcaattaac tgtacaagac ccaacaacaa tacaagaaaa agtatctata
     2581 taggaccagg gagagcattt catacaacag gaagaataat aggagatata agaaaagcac
     2641 attgtaacat tagtagagca caatggaata acactttaga acagatagtt aaaaaattaa
     2701 gagaacagtt tgggaataat aaaacaatag tctttaatca atcctcagga ggggacccag
     2761 aaattgtaat gcacagtttt aattgtagag gggaattttt ctactgtaat acaacacaac
     2821 tgtttaataa tacatggagg ttaaatcaca ctgaaggaac taaaggaaat gacacaatca
     2881 tactcccatg tagaataaaa caaattataa acatgtggca ggaagtagga aaagcaatgt
     2941 atgcccctcc cattggagga caaattagtt gttcatcaaa tattacaggg ctgctattaa
     3001 caagagatgg tggtacaaat gtaactaatg acaccgaggt cttcagacct ggaggaggag
     3061 atatgaggga caattggaga agtgaattat ataaatataa agtaataaaa attgaaccat
     3121 taggaatagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     3181 gaatagtagg agctatgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     3241 tgtcattgac gctgacggta caggccagac aattattgtc tggtatagtg caacagcaga
     3301 acaatttgct gagggctatt gaggcgcaac aacatctgtt gcaactcaca gtctggggca
     3361 tcaagcagct ccaggcaaga gtcctggctg tggaaagata cctaagggat caacagctcc
     3421 tagggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     3481 gttggagtaa taaatctctg gaagacattt gggataacat gacctggatg cagtgggaaa
     3541 gagaaattga caattacaca aacacaatat acaccttact tgaagaatcg cagaaccaac
     3601 aagaaaagaa tgaacaagaa ttattagaat tggataagtg ggcaagtttg tggaattggt
     3661 ttagcataac aaactggctg tggtatataa agatattcat aatgatagta ggaggcttgg
     3721 taggtttaag aatagttttt gctgtgcttt ctatagtgaa tagagttagg cagggatact
     3781 caccattgtc atttcagacc cgcctcccag tctcgagacc tggaaaaaca tggagcaatc
     3841 acaagtagca atacagcagc tactaatgct gattgtgcct ggctagaagc acaagaggag
     3901 gaagaggtgg gttttccagt cagacctcag gtacctttaa gaccaatgac ttacaaggca
     3961 gctttagata ttagccactt tttaaaagaa aaggggggac tggaagggct aatttggtcc
     4021 caaagaagac aagagagatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     4081 attggcagaa ttacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct
     4141 tcaagctagt accagttgag ccagagaagg tagaagaggc caatgaagga gagaacaaca
     4201 gcttgttaca ccctatgagc ctgcatggga tggaggacgc ggagaaagaa gtgttagtgt
     4261 ggaggtttga cagcaaacta gcatttcatc acatggcccg agagctgcat ccggagtact
     4321 acaaagactg ctgacatcga gctttctaca agggactttc cgctggggac tttccaggga
     4381 ggcgtggcct gggcgggact ggggagtggc gtgccctcag atgctgcata taagcagctg
     4441 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     4501 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     4561 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4621 tggaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4681 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4741 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     4801 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     4861 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4921 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4981 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5041 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5101 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5161 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5221 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5281 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5341 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5401 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5461 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5521 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5581 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5641 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5701 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5761 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5821 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5881 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5941 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6001 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6061 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6121 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6181 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6361 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6421 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6481 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6541 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6601 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6661 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6721 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6781 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6841 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6901 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6961 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7021 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7081 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7141 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7201 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7261 ctt
//
