LOCUS       sequence-503503-        6243 bp    DNA     circular SYN 19-MAR-2026
DEFINITION  Plasmid for studying nuclear export. Expression of a fusion protein 
            containing full-length HIV-1 Rev, followed by the hormone-responsive
            element of the rat glucocorticoid receptor (Gr), and GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6243)
  AUTHORS   Takao D, Dishinger JF, Kee HL, Pinskey JM, Allen BL, Verhey KJ
  TITLE     An assay for clogging the ciliary pore complex distinguishes 
            mechanisms of cytosolic and membrane protein entry.
  JOURNAL   Curr Biol. 2014 Oct 6;24(19):2288-94. doi: 
            10.1016/j.cub.2014.08.012. Epub 2014 Sep 25.
  PUBMED    25264252
REFERENCE   2  (bases 1 to 6243)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 19, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cub.2014.08.012"; journalName: "Curr Biol"; date: 
            "2014-10-6- 6"; volume: "24"; issue: "19"; pages: "2288-94"
COMMENT     Sequence Label: pN1_Rev-GR-EGFP
FEATURES             Location/Qualifiers
     source          1..6243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(834..938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1006..1024
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1051..1056,1057..1074,1075..1081)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1089..1105
                     /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)."
     enhancer        1135..1438
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1439..1642
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1593..1613
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1639..1663
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1758..1776
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1758..1775
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1856..2200
                     /codon_start=1
                     /gene="rev"
                     /product="human immunodeficiency virus 1 (HIV-1) Rev 
                     protein"
                     /label=Rev
                     /note="enhances nuclear export of unspliced and 
                     incompletely spliced viral mRNAs"
                     /translation="MAGRSGDSDEDLLKAVRLIKFLYQSNPPPNPEGTRQARRNRRRRW
                     RERQRQIHSISERILSTYLGRSAEPVPLQLPPLERLTLDCNEDCGTSGTQGVGSPQILV
                     ESPTILESGAK"
     polyA_signal    4170..4304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4207..4226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4261..4280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4430..4884
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4517..4536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4726..4747
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(5171..5239,5240..6031)
                     /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(5389..5408)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    6133..6166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagagctt gcaattcgcg
      841 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      901 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      961 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1021 aggccctttc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     1081 gtgtggaatt gtgagcggat aacaatttca cataggaaac agcatcgtgc aggtcgttac
     1141 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     1201 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     1261 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     1321 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     1381 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     1441 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1501 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1561 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1621 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
     1681 acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga ctctagccta
     1741 ggcttttgca aaaagctatt taggtgacac tatagaaggt acgcctgcag gtaccggtcc
     1801 ggaattcccg ggtcgacgag ctcactagtc ggcggccgcc gccaccatgc tcgagatggc
     1861 aggaagaagc ggagacagcg acgaagacct cctcaaggca gtcagactca tcaagtttct
     1921 ctatcaaagc aacccacctc ccaatcccga ggggacccga caggcccgaa ggaatagaag
     1981 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tccttagcac
     2041 ttatctggga cgatctgcgg agcctgtgcc tcttcagcta ccaccgcttg agagacttac
     2101 tcttgattgt aacgaggatt gtggaacttc tgggacgcag ggggtgggaa gccctcaaat
     2161 attggtggaa tctcctacaa tattggagtc aggagctaaa gatctgacaa agaaaaaaat
     2221 caaagggatt cagcaagcca ctgcaggagt ctcacaagac acttcggaaa atcctaacaa
     2281 aacaatagtt cctgctgcat taccacagct cacccctacc ttggtgtcac tgctggaggt
     2341 gattgaaccc gaggtgttgt atgcaggata tgatagctct gttccagatt cagcatggag
     2401 aattatgacc acactcaaca tgttaggtgg gcgtcaagtg attgcagcag tgaaatgggc
     2461 aaaggcgata ccaggcttca gaaacttaca cctggatgac caaatgaccc tgctacagta
     2521 ctcatggatg tttctcatgg catttgccct gggttggaga tcatacagac aatcaagtgg
     2581 aaacctgctc tgctttgctc ctgatctgat tattaatgag cagagaatgt ctctaccctg
     2641 catgtatgac caatgtaaac acatgctgtt tgtctcctct gaattacaaa gattgcaggt
     2701 atcctatgaa gagtatctct gtatgaaaac cttactgctt ctctcctcag ttcctaagga
     2761 aggtctgaag agccaagagt tatttgatga gattcgaatg acttatatca aagagctagg
     2821 aaaagccatc gtcaaaaggg aagggaactc cagtcagaac tggcaacggt tttaccaact
     2881 gacaaagctt ctggactcca tgcatgaggt ggttgagaat ctccttacct actgcttcca
     2941 gacatttttg gataagacca tgagtattga attcccagag atgttagctg aaatcatcac
     3001 taatcagata ccaaaatatt caaatggaaa tatcaaaaag cttctgtttc atcaaaaact
     3061 cgagagcaag ggcgaggaac tgttcactgg cgtggtccca attctcgtgg aactggatgg
     3121 cgatgtgaat gggcacaaat tttctgtcag cggagagggt gaaggtgatg ccacatacgg
     3181 aaagctcacc ctgaaattca tctgcaccac tggaaagctc cctgtgccat ggccaacact
     3241 ggtcactacc ttcacctatg gcgtgcagtg cttttccaga tacccagacc atatgaagca
     3301 gcatgacttt ttcaagagcg ccatgcccga gggctatgtg caggagagaa ccatcttttt
     3361 caaagatgac gggaactaca agacccgcgc tgaagtcaag ttcgaaggtg acaccctggt
     3421 gaatagaatc gagctgaagg gcattgactt taaggaggat ggaaacattc tcggccacaa
     3481 gctggaatac aactataact cccacaatgt gtacatcatg gccgacaagc aaaagaatgg
     3541 catcaaggtc aacttcaaga tcagacacaa cattgaggat ggatccgtgc agctggccga
     3601 ccattatcaa cagaacactc caatcggcga cggccctgtg ctcctcccag acaaccatta
     3661 cctgtccacc cagtctgccc tgtctaaaga tcccaacgaa aagagagacc acatggtcct
     3721 gctggagttt gtgaccgctg ctgggatcac acatggcatg gacgagctgt acaagtgagc
     3781 ggccgctcta gcttgggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
     3841 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
     3901 gtgttaaact agctgcatat gcttgctgct tgagagtttt gcttactgag tatgatttat
     3961 gaaaatatta tacacaggag ctagtgattc taattgtttg tgtattttag attcacagtc
     4021 ccaaggctca tttcaggccc ctcagtcctc acagtctgtt catgatcata atcagccata
     4081 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4141 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4201 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4261 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcgatcct gcattaatga
     4321 atcggccaac gcgcggggag aggcggtttg cgtattggct ggcgtaatag cgaagaggcc
     4381 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     4441 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4501 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4561 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4621 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4681 tagacggttt ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc
     4741 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4801 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4861 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4921 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgccag gtcttggact
     4981 ggtgagaacg gcttgctcgg cagcttcgat gtgtgctgga gggagaataa aggtctaaga
     5041 tgtgcgatag agggaagtcg cattgaatta tgtgctgtgt agggatcgct ggtatcaaat
     5101 atgtgtgccc acccctggca tgagacaata accctgataa atgcttcaat aatattgaaa
     5161 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5221 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5281 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5341 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5401 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5461 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5521 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5581 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5641 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5701 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5761 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5821 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5881 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5941 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6001 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6061 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6121 taatctcatg ccataacttc gtataatgta tgctatacga agttatggca tgaccaaaat
     6181 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6241 ttc
//
