LOCUS       sequence-289555-        7218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Rluc-CTE-GAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7218)
  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 7218)
  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-CTE-GAG_O
FEATURES             Location/Qualifiers
     source          1..7218
                     /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..4242
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             5803..5847
                     /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(6824..6841)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6824..6840)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6833..6855)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7049..7068)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7168..7190
                     /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 cttccaaggt gtacgacccc gagcaacgca aacgcatgat cactgggcct
     3361 cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa ctactatgat
     3421 tccgagaagc acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc ctccagctac
     3481 ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat ccctgatctg
     3541 atcggaatgg gtaagtccgg caagagcggg aatggctcat atcgcctcct ggatcactac
     3601 aagtacctca ccgcttggtt cgagctgctg aaccttccaa agaaaatcat ctttgtgggc
     3661 cacgactggg gggcttgtct ggcctttcac tactcctacg agcaccaaga caagatcaag
     3721 gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt cctgggacga gtggcctgac
     3781 atcgaggagg atatcgccct gatcaagagc gaagagggcg agaaaatggt gcttgagaat
     3841 aacttcttcg tcgagaccat gctcccaagc aagatcatgc ggaaactgga gcctgaggag
     3901 ttcgctgcct acctggagcc attcaaggag aagggcgagg ttagacggcc taccctctcc
     3961 tggcctcgcg agatccctct cgttaaggga ggcaagcccg acgtcgtcca gattgtccgc
     4021 aactacaacg cctaccttcg ggccagcgac gatctgccta agatgttcat cgagtccgac
     4081 cctgggttct tttccaacgc tattgtcgag ggagctaaga agttccctaa caccgagttc
     4141 gtgaaggtga agggcctcca cttcagccag gaggacgctc cagatgaaat gggtaagtac
     4201 atcaagagct tcgtggagcg cgtgctgaag aacgagcagt aaggatcctc tagtctgtgt
     4261 tactttaaat tgcactgatt tggggaaggc tactaatacc aatagtagta attggaaaga
     4321 agaaataaaa ggagaaataa aaaactgctc tttcaatatc accacaagca taagagataa
     4381 gattcagaaa gaaaatgcac tttttcgtaa ccttgatgta gtaccaatag ataatgctag
     4441 tactactacc aactatacca actataggtt gatacattgt aacagatcag tcattacaca
     4501 ggcctgtcca aaggtatcat ttgagccaat tcccatacat tattgtaccc cggctggttt
     4561 tgcgattcta aagtgtaata ataaaacgtt caatggaaaa ggaccatgta caaatgtcag
     4621 cacagtacaa tgtacacatg gaattaggcc aatagtgtca actcaactgc tgttaaatgg
     4681 cagtctagca gaagaagagg tagtaattag atctgacaat ttcacgaaca atgctaaaac
     4741 cataatagta cagctgaatg aatctgtagc aattaactgt acaagaccca acaacaatac
     4801 aagaaaaagt atctatatag gaccagggag agcatttcat acaacaggaa gaataatagg
     4861 agatataaga aaagcacatt gtaacattag tagagcacaa tggaataaca ctttagaaca
     4921 gatagttaaa aaattaagag aacagtttgg gaataataaa acaatagtct ttaatcaatc
     4981 ctcaggaggg gacccagaaa ttgtaatgca cagttttaat tgtagagggg aatttttcta
     5041 ctgtaataca acacaactgt ttaataatac atggaggtta aatcacactg aaggaactaa
     5101 aggaaatgac acaatcatac tcccatgtag aataaaacaa attataaaca tgtggcagga
     5161 agtaggaaaa gcaatgtatg cccctcccat tggaggacaa attagttgtt catcaaatat
     5221 tacagggctg ctattaacaa gagatggtgg tacaaatgta actaatgaca ccgaggtctt
     5281 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     5341 aataaaaatt gaaccattag gaatagcacc caccaaggca aagagaagag tggtgcagag
     5401 agaaaaaaga gcagtgggaa tagtaggagc tatggggccc agaccacctc ccctgcgagc
     5461 taagctggac agccaatgac gggtaagaga gtgacatttt tcactaacct agagcaggag
     5521 ggccgtcaga gctactgcct aatccaaaga cgggtaaaag tgataaaaat gtatcactcc
     5581 aacctagagc aggcgcagct tccgagggat tcccgggtag ggatttgggg ttgctctgga
     5641 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     5701 gacatttggg ataacatgac ctggatgcag tgggaaagag aaattgacaa ttacacaaac
     5761 acaatataca ccttacttga agaatcgcag aaccaacaag aaaagaatga acaagaatta
     5821 ttagaattgg ataagtgggc aagtttgtgg aattggttta gcataacaaa ctggctgtgg
     5881 tatataaaga tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     5941 gtgctttcta tagtgaatag agttaggcag ggatactcac cattgtcatt tcagacccgc
     6001 ctcccagtcc tcgagacctg gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     6061 ctaatgctga ttgtgcctgg ctagaagcac aagaggagga agaggtgggt tttccagtca
     6121 gacctcaggt acctttaaga ccaatgactt acaaggcagc tttagatatt agccactttt
     6181 taaaagaaaa ggggggactg gaagggctaa tttggtccca aagaagacaa gagatccttg
     6241 atctgtggat ctaccacaca caaggctact tccctgattg gcagaattac acaccagggc
     6301 cagggatcag atatccactg acctttggat ggtgcttcaa gctagtacca gttgagccag
     6361 agaaggtaga agaggccaat gaaggagaga acaacagctt gttacaccct atgagcctgc
     6421 atgggatgga ggacgcggag aaagaagtgt tagtgtggag gtttgacagc aaactagcat
     6481 ttcatcacat ggcccgagag ctgcatccgg agtactacaa agactgctga catcgagctt
     6541 tctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg
     6601 agtggcgtcc ctcagatgct gcatataagc agctgctttt tgcctgtact gggtctctct
     6661 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6721 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6781 gtaactagag atccctcaga cccttttagt cagtgtggaa ttcactggcc gtcgttttac
     6841 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6901 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6961 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     7021 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7081 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     7141 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     7201 catcaccgaa acgcgcga
//
