LOCUS       sequence-418111-        5919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible AAV activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5919)
  AUTHORS   Tycko J, Van MV, Aradhana, DelRosso N, Ye H, Yao D, Valbuena R, 
            Vaughan-Jackson A, Xu X, Ludwig C, Spees K, Liu K, Gu M, Khare V, 
            Mukund AX, Suzuki PH, Arana S, Zhang C, Du PP, Ornstein TS, Hess GT,
            Kamber RA, Qi LS, Khalil AS, Bintu L, Bassik MC
  TITLE     Development of compact transcriptional effectors using 
            high-throughput measurements in diverse contexts.
  JOURNAL   Nat Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6.
  PUBMED    39487265
REFERENCE   2  (bases 1 to 5919)
  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"; journalName: "Nat 
            Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6."
COMMENT     Sequence Label: pJT410 AAV-Rapamycin Inducible-NFZ-Citrine wit...
FEATURES             Location/Qualifiers
     source          1..5919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..152
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   23..152
                     /label=AAV2 ITR
     protein_bind    159..179
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             334..360
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1090..1143
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1144..1170
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             join(1177..1350,1351..1356,1357..1539)
                     /codon_start=1
                     /product="composite human DNA-binding domain composed of 
                     domains from the transcription factors Zif268 and Oct-1 
                     (Pomerantz et al., 1995)"
                     /label=ZFHD1
                     /label=ZFHD1 (DNA binding domain)
                     /translation="ERPYACPVESCDRRFSRSDELTRHIRIHTGQKPFQCRICMRNFSR
                     SDHLTTHIRTHTGGGRRRKKRTSIETNIRVALEKSFLENQKPTSEEITMIADQLNMEKE
                     VIRVWFCNRRQKEKRIN"
     CDS             1546..1869
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1546..1866
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    1876..2083
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2262..2375
                     /label=minimal IL-2 promoter
                     /label=IL-2 promoter
                     /note="minimal promoter from the human interleukin-2 gene"
     regulatory      2379..2388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2385..2387,2388..2390,2391..3101)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2430..2451)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2691..2710)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3038..3059
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   3193..3333
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3193..3322
                     /label=AAV2 ITR
     rep_origin      3408..3863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3495..3514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3705..3726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3880..3899)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3999..4021
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4059..4077)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4145..4249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4250..4318,4319..5110)
                     /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(4468..4487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5281..5869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5770..5789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
       61 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
      121 gggagtggcc aactccatca ctaggggttc cttctagaca actttgtata gaaaagttgg
      181 gtaccgcggc cgcacgcgtt aatgatgggc gctacagtaa tgatgggcgg aattcctcga
      241 ggtacttata taagggggtg ggggcgcgtt cgtcctcagt cgcgatcgaa cactcgagcc
      301 gagcagacgt gcctaaccgg tgccgccacc atgcctgctg ccaagagggt caagttggac
      361 tggcatgaag gcctggaaga ggcatctcgt ttgtactttg gggaaaggaa cgtgaaaggc
      421 atgtttgagg tgctggagcc cttgcatgct atgatggaac ggggccccca gactctgaag
      481 gaaacatctt ttaatcaggc ctatggtcgt gatttaatgg aggcccaaga gtggtgcagg
      541 aagtacatga aatcagggaa tgtcaaggac ctcacccaag cctgggacct ctattatcat
      601 gtgttccgac gaatctcaaa gcagtctgga tccggatcag gctccgaggg ccagagcgac
      661 gagagagccc tgctggacca gctgcacacc ctgctgagca acaccgacgc caccggcctg
      721 gaggagatcg acagagccct gggcatcccc gagctggtga accagggcca ggccctggag
      781 cccaagcagg gaggaggagg tagcggacac gagaagttcc ccagcgacct ggacctggac
      841 atgttcaacg gcagcctgga gtgcgacatg gagagcatca tcagaagcga gctgatggac
      901 gccgacggcc tggacttcaa cttcgacagc ggaggaggag gtagcggatt cgtgaccctg
      961 aaggacgtgg gcatggactt caccctgggc gactgggagc agctgggcct ggagcagggc
     1021 gacaccttct gggacaccgc cctggacaac tgccaggacc tgttcctgct ggaccccccc
     1081 ggatctggag agggaagggg aagcctccta acttgcggag atgtcgagga gaatcctggc
     1141 ccacctgccg ctaagcgtgt caagttggac tctagagaac gcccgtatgc ttgccctgtc
     1201 gagtcctgcg atcgccgctt ttctcgctcg gatgagctta cccgccatat ccgcatccac
     1261 acaggccaga agcccttcca gtgtcgaatc tgcatgcgta acttcagtcg tagtgaccac
     1321 cttaccaccc acatccgcac ccacacaggc ggcggccgca ggaggaagaa acgcaccagc
     1381 atagagacta acatccgtgt ggccttagag aagagtttct tggagaatca aaagcctacc
     1441 tcggaagaga tcactatgat tgctgatcag ctcaatatgg aaaaagaggt gattcgtgtt
     1501 tggttctgta accgccgcca gaaagaaaaa agaatcaaca agggcggcgt gcaggtggag
     1561 actatctccc caggcgacgg gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac
     1621 tacaccggga tgcttgaaga tggaaagaaa tttgattcct cccgggacag aaacaagccc
     1681 tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag
     1741 atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta tggtgccact
     1801 gggcacccag gcatcatccc accacatgcc actctcgtct ttgatgtgga gcttctaaaa
     1861 ctggaataac atatgctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     1921 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1981 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2041 gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggacgtacgt aatgatgggc
     2101 gctaaattaa tgatgggcgg agttgtaatg atgggcgctc gtttaatgat gggcgtctag
     2161 ctaatgatgg gcgggcgatt aatgatgggc gaaggactaa tgatgggcgc agcggtaatg
     2221 atgggcgatc agctaatgat gggcgacgcg aattaattca acattttgac acccccataa
     2281 tatttttcca gaattaacag tataaattgc atctcttgtt caagagttcc ctatcactct
     2341 ctttaatcac tactcacagt aacctcaact cctgcgccgc caccatggtg agcaagggcg
     2401 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2461 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2521 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
     2581 gctacggcct gatgtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2641 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2701 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2761 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2821 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2881 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2941 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3001 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3061 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagcctaa taaagagctc
     3121 agatgcatgc atcagagtgt gttggttttt tgtgtgtcgg accgagcggc cgcgcatgct
     3181 ggggagggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3241 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3301 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3361 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3421 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3481 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3541 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3601 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3661 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3721 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3781 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3841 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3901 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3961 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4021 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4081 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4141 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4201 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4261 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4321 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4381 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4441 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4501 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4561 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4621 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4681 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4741 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4801 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4861 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4921 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     4981 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5041 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5101 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5161 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5221 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5281 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5341 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5401 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5461 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5521 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5581 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5641 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5701 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5761 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5821 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5881 cgcggccttt ttacggttcc tggccttttg ctggccttt
//
