LOCUS       sequence-289551-        7108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of CAT-RRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7108)
  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 7108)
  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-CAT-RRE_O
FEATURES             Location/Qualifiers
     source          1..7108
                     /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"
     CDS             3345..4004
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3411..3430)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     misc_feature    5275..5508
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5693..5737
                     /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(6714..6731)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6714..6730)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6723..6745)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6939..6958)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7058..7080
                     /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 cggccgcaag
     3301 gggatctgag cttggcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     3361 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     3421 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     3481 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     3541 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
     3601 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     3661 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     3721 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     3781 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     3841 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     3901 cgctggcgat tcaggttcat catgccgtct gtgatggctt ccatgtcggc agaatgctta
     3961 atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt
     4021 attggtgccc ttaaacgcct ggtgctacgc ctgaataagt gataataagc ggatgaatgg
     4081 cagaaattcg ccggatcctc tagtctgtgt tactttaaat tgcactgatt tggggaaggc
     4141 tactaatacc aatagtagta attggaaaga agaaataaaa ggagaaataa aaaactgctc
     4201 tttcaatatc accacaagca taagagataa gattcagaaa gaaaatgcac tttttcgtaa
     4261 ccttgatgta gtaccaatag ataatgctag tactactacc aactatacca actataggtt
     4321 gatacattgt aacagatcag tcattacaca ggcctgtcca aaggtatcat ttgagccaat
     4381 tcccatacat tattgtaccc cggctggttt tgcgattcta aagtgtaata ataaaacgtt
     4441 caatggaaaa ggaccatgta caaatgtcag cacagtacaa tgtacacatg gaattaggcc
     4501 aatagtgtca actcaactgc tgttaaatgg cagtctagca gaagaagagg tagtaattag
     4561 atctgacaat ttcacgaaca atgctaaaac cataatagta cagctgaatg aatctgtagc
     4621 aattaactgt acaagaccca acaacaatac aagaaaaagt atctatatag gaccagggag
     4681 agcatttcat acaacaggaa gaataatagg agatataaga aaagcacatt gtaacattag
     4741 tagagcacaa tggaataaca ctttagaaca gatagttaaa aaattaagag aacagtttgg
     4801 gaataataaa acaatagtct ttaatcaatc ctcaggaggg gacccagaaa ttgtaatgca
     4861 cagttttaat tgtagagggg aatttttcta ctgtaataca acacaactgt ttaataatac
     4921 atggaggtta aatcacactg aaggaactaa aggaaatgac acaatcatac tcccatgtag
     4981 aataaaacaa attataaaca tgtggcagga agtaggaaaa gcaatgtatg cccctcccat
     5041 tggaggacaa attagttgtt catcaaatat tacagggctg ctattaacaa gagatggtgg
     5101 tacaaatgta actaatgaca ccgaggtctt cagacctgga ggaggagata tgagggacaa
     5161 ttggagaagt gaattatata aatataaagt aataaaaatt gaaccattag gaatagcacc
     5221 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa tagtaggagc
     5281 tatgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagtgt cattgacgct
     5341 gacggtacag gccagacaat tattgtctgg tatagtgcaa cagcagaaca atttgctgag
     5401 ggctattgag gcgcaacaac atctgttgca actcacagtc tggggcatca agcagctcca
     5461 ggcaagagtc ctggctgtgg aaagatacct aagggatcaa cagctcctag ggatttgggg
     5521 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     5581 atctctggaa gacatttggg ataacatgac ctggatgcag tgggaaagag aaattgacaa
     5641 ttacacaaac acaatataca ccttacttga agaatcgcag aaccaacaag aaaagaatga
     5701 acaagaatta ttagaattgg ataagtgggc aagtttgtgg aattggttta gcataacaaa
     5761 ctggctgtgg tatataaaga tattcataat gatagtagga ggcttggtag gtttaagaat
     5821 agtttttgct gtgctttcta tagtgaatag agttaggcag ggatactcac cattgtcatt
     5881 tcagacccgc ctcccagtcc tcgagacctg gaaaaacatg gagcaatcac aagtagcaat
     5941 acagcagcta ctaatgctga ttgtgcctgg ctagaagcac aagaggagga agaggtgggt
     6001 tttccagtca gacctcaggt acctttaaga ccaatgactt acaaggcagc tttagatatt
     6061 agccactttt taaaagaaaa ggggggactg gaagggctaa tttggtccca aagaagacaa
     6121 gagatccttg atctgtggat ctaccacaca caaggctact tccctgattg gcagaattac
     6181 acaccagggc cagggatcag atatccactg acctttggat ggtgcttcaa gctagtacca
     6241 gttgagccag agaaggtaga agaggccaat gaaggagaga acaacagctt gttacaccct
     6301 atgagcctgc atgggatgga ggacgcggag aaagaagtgt tagtgtggag gtttgacagc
     6361 aaactagcat ttcatcacat ggcccgagag ctgcatccgg agtactacaa agactgctga
     6421 catcgagctt tctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc
     6481 gggactgggg agtggcgtcc ctcagatgct gcatataagc agctgctttt tgcctgtact
     6541 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6601 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6661 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa ttcactggcc
     6721 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     6781 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     6841 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     6901 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     6961 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     7021 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     7081 ttttcaccgt catcaccgaa acgcgcga
//
