LOCUS       sequence-409995-        7223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Rluc-CTE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7223)
  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 7223)
  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_O
FEATURES             Location/Qualifiers
     source          1..7223
                     /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"
     CDS             3566..3610
                     /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(4589..4606)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4589..4605)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4598..4620)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4814..4833)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4933..4955
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4993..5011)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5079..5183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5184..5252,5253..6044)
                     /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(5402..5421)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6215..6803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6704..6723
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6957..6974
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7091..7112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7127..7132,7133..7150,7151..7157)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7165..7181
                     /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     7170..7192
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7189..7205
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7189..7205
                     /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 agctatgggg cccagaccac ctcccctgcg agctaagctg gacagccaat
     3241 gacgggtaag agagtgacat ttttcactaa cctaagacag gagggccgtc agagctactg
     3301 cctaatccaa agacgggtaa aagtgataaa aatgtatcac tccaacctaa gacaggcgca
     3361 gcttccgagg gattcccggg tagggatttg gggttgctct ggaaaactca tttgcaccac
     3421 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaagacattt gggataacat
     3481 gacctggatg cagtgggaaa gagaaattga caattacaca aacacaatat acaccttact
     3541 tgaagaatcg cagaaccaac aagaaaagaa tgaacaagaa ttattagaat tggataagtg
     3601 ggcaagtttg tggaattggt ttagcataac aaactggctg tggtatataa agatattcat
     3661 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtgcttt ctatagtgaa
     3721 tagagttagg cagggatact caccattgtc atttcagacc cgcctcccag tctcgagacc
     3781 tggaaaaaca tggagcaatc acaagtagca atacagcagc tactaatgct gattgtgcct
     3841 ggctagaagc acaagaggag gaagaggtgg gttttccagt cagacctcag gtacctttaa
     3901 gaccaatgac ttacaaggca gctttagata ttagccactt tttaaaagaa aaggggggac
     3961 tggaagggct aatttggtcc caaagaagac aagagagatc cttgatctgt ggatctacca
     4021 cacacaaggc tacttccctg attggcagaa ttacacacca gggccaggga tcagatatcc
     4081 actgaccttt ggatggtgct tcaagctagt accagttgag ccagagaagg tagaagaggc
     4141 caatgaagga gagaacaaca gcttgttaca ccctatgagc ctgcatggga tggaggacgc
     4201 ggagaaagaa gtgttagtgt ggaggtttga cagcaaacta gcatttcatc acatggcccg
     4261 agagctgcat ccggagtact acaaagactg ctgacatcga gctttctaca agggactttc
     4321 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gtgccctcag
     4381 atgctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     4441 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     4501 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     4561 tcagaccctt ttagtcagtg tggaattcac tggccgtcgt tttacaacgt cgtgactggg
     4621 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4681 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4741 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     4801 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4861 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4921 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4981 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5041 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5101 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5161 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5221 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5281 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5341 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5401 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5461 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5521 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5581 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5641 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5701 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5761 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5821 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5881 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5941 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6001 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6061 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6121 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6181 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6241 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6301 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6361 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6421 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6481 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6541 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6601 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6661 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6721 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6781 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6841 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6901 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6961 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     7021 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     7081 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     7141 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     7201 atgaccatga ttacgccaag ctt
//
