LOCUS       sequence-423199-        8038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses one of the split transcription factor component in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8038)
  AUTHORS   Shen J, Geng L, Li X, Emery C, Kroning K, Shingles G, Lee K, Heyden 
            M, Li P, Wang W
  TITLE     A general method for chemogenetic control of peptide function.
  JOURNAL   Nat Methods. 2023 Jan;20(1):112-122. doi: 
            10.1038/s41592-022-01697-8. Epub 2022 Dec 8.
  PUBMED    36481965
REFERENCE   2  (bases 1 to 8038)
  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/s41592-022-01697-8"; journalName: "Nat Methods"; date: 
            "2023-01"; volume: "20"; issue: "1"; pages: "112-122"
COMMENT     Sequence Label: plx208-CMV-Gal4DBD-CapN-SsrA-CapC
FEATURES             Location/Qualifiers
     source          1..8038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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"
     misc_feature    1847..1964
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2081..2384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2385..2588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2539..2559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2617..2640
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2641..3078
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3079..3099
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3556..3879
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    3895..4483
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3948..3968)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4366..4377)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4555..4788
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4866..4987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4903..4922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4957..4976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5027..5162
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5182..5201)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5183..5201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5211..5228)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5211..5227)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5220..5242)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5369..5824
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5456..5475)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5666..5687
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5850..5954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5955..6023,6024..6815)
                     /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(6173..6192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6986..7574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7475..7494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7728..7745
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7862..7883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7898..7903,7904..7921,7922..7928)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7936..7952
                     /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     7941..7963
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7960..7976
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7960..7976
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7995..8015
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7997..8015
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagagcg ctgccgtctg gaccgatggc tgtgtggaag
     1741 tgctcgccga tagtggaaac agacgcccca gcactcgtcc tagggcaaag gatctgcagt
     1801 aatgagaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     1861 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     1921 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     1981 acagcagaga tccactttgg cgccggctcg agggggcccg gggttattaa tagtaatcaa
     2041 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2101 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2161 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2221 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2281 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2341 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2401 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2461 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2521 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2581 gcagagctct ctggctaact gtcgggatcc accatggact acaaggatga cgacgataag
     2641 aagctactgt cttctatcga acaagcatgc gatatttgcc gacttaaaaa gctcaagtgc
     2701 tccaaagaaa aaccgaagtg cgccaagtgt ctgaagaaca actgggagtg tcgctactct
     2761 cccaaaacca aaaggtctcc gctgactagg gcacatctga cagaagtgga atcaaggcta
     2821 gaaagactgg aacagctatt tctactgatt tttcctcgag aagaccttga catgattttg
     2881 aaaatggatt ctttacagga tataaaagca ttgttaacag gattatttgt acaagataat
     2941 gtgaataaag atgccgtcac agatagattg gcttcagtgg agactgatat gcctctaaca
     3001 ttgagacagc atagaataag tgcgacatca tcatcggaag agagtagtaa caaaggtcaa
     3061 agacagttga ctgtatcgcc caagaagaag aggaaggtgg ctagcggcgt ccaagtcgag
     3121 acaatatctc ccggcgacgg acgaacgttt ccgaagcggg gacagacttg cgtcgttcac
     3181 tacacaggaa tgctggagga tggcaaaaag gtcgattcca gccgggacag gaataagccg
     3241 ttcaaattca tgctgggaaa acaggaggta atacgaggtt gggaggaggg tgtggcgcaa
     3301 atgtctgtcg gtcagcgggc gaaactgacc atttctcctg attatgctta cggagcgaca
     3361 ggccatccgg ggatcattcc gccccacgct accctggtat tcgatgtaga actcttgaag
     3421 ctcgaaacca gaggggttga agaagtagcc agatattccc ccaacctggc agcgaatgat
     3481 gaaaattacg gtactcctaa tttgcggcct tttggttcag gcggttctgg tactggatct
     3541 ggttctggag gttctggagt gcaggtggaa accatctccc caggagacgg gcgcaccttc
     3601 cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa
     3661 gttgattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa gcaggaggtg
     3721 atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc caaactgact
     3781 atatctccag attatgccta tggtgccact gggcacccag gcatcatccc accacatgcc
     3841 actctcgtct tcgatgtgga gcttctaaaa ctggaataga cgcgttaagt cgacaatcaa
     3901 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3961 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4021 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4081 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4141 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4201 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4261 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4321 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4381 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4441 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
     4501 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     4561 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     4621 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     4681 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     4741 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta
     4801 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     4861 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4921 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4981 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     5041 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5101 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5161 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     5221 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     5281 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5341 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     5401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5521 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5581 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5701 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5761 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     5821 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5881 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5941 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6001 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6061 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6121 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6181 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6241 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6301 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6361 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6421 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6481 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6541 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6601 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6661 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6721 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6781 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6841 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6901 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6961 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7021 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7081 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7141 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7201 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7261 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7321 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7381 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7441 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7501 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7561 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7621 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7681 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7741 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     7801 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     7861 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7921 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     7981 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagc
//
