LOCUS       sequence-418108-        5491 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 5491)
  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 5491)
  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: pJT313 AAV-Rapamycin Inducible-ZF-Citrine
FEATURES             Location/Qualifiers
     source          1..5491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(19..87,88..879)
                     /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(237..256)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1050..1638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1539..1558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1700..1829
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1978..2004
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2572..2625
                     /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             2626..2652
                     /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(2659..2832,2833..2838,2839..3021)
                     /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             3028..3351
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3028..3348
                     /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"
     promoter        3553..3666
                     /label=minimal IL-2 promoter
                     /label=IL-2 promoter
                     /note="minimal promoter from the human interleukin-2 gene"
     regulatory      3670..3679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3676..3678,3679..3681,3682..4392)
                     /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(3721..3742)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3982..4001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4329..4350
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   4465..4605
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4465..4594
                     /label=AAV2 ITR
     rep_origin      4680..5135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4767..4786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4977..4998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5152..5171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5265..5287
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5319..5337)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5405..5491,1..18)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
       61 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      121 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
      181 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
      241 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
      301 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
      361 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
      421 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
      481 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
      541 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
      601 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
      661 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
      721 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
      781 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
      841 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
      901 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
      961 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1021 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1141 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1201 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1261 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1321 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1381 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1441 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1501 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1561 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1621 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1681 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     1741 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     1801 agtggccaac tccatcacta ggggttcctg cggccgcacg cgttaatgat gggcgctaca
     1861 gtaatgatgg gcggaattcc tcgaggtact tatataaggg ggtgggggcg cgttcgtcct
     1921 cagtcgcgat cgaacactcg agccgagcag acgtgcctaa ccggtgccgc caccatgcct
     1981 gctgccaaga gggtcaagtt ggactggcat gaaggcctgg aagaggcatc tcgtttgtac
     2041 tttggggaaa ggaacgtgaa aggcatgttt gaggtgctgg agcccttgca tgctatgatg
     2101 gaacggggcc cccagactct gaaggaaaca tcttttaatc aggcctatgg tcgtgattta
     2161 atggaggccc aagagtggtg caggaagtac atgaaatcag ggaatgtcaa ggacctcacc
     2221 caagcctggg acctctatta tcatgtgttc cgacgaatct caaagcagtc tggatccgga
     2281 tcaggctcct tcgtgaccct gaaggacgtg ggcatggact tcaccctggg cgactgggag
     2341 cagctgggcc tggagcaggg cgacaccttc tgggacaccg ccctggacaa ctgccaggac
     2401 ctgttcctgc tggacccccc cggaggagga ggtagcggac acgagaagtt ccccagcgac
     2461 ctggacctgg acatgttcaa cggcagcctg gagtgcgaca tggagagcat catcagaagc
     2521 gagctgatgg acgccgacgg cctggacttc aacttcgaca gcggatctgg agagggaagg
     2581 ggaagcctcc taacttgcgg agatgtcgag gagaatcctg gcccacctgc cgctaagcgt
     2641 gtcaagttgg actctagaga acgcccgtat gcttgccctg tcgagtcctg cgatcgccgc
     2701 ttttctcgct cggatgagct tacccgccat atccgcatcc acacaggcca gaagcccttc
     2761 cagtgtcgaa tctgcatgcg taacttcagt cgtagtgacc accttaccac ccacatccgc
     2821 acccacacag gcggcggccg caggaggaag aaacgcacca gcatagagac taacatccgt
     2881 gtggccttag agaagagttt cttggagaat caaaagccta cctcggaaga gatcactatg
     2941 attgctgatc agctcaatat ggaaaaagag gtgattcgtg tttggttctg taaccgccgc
     3001 cagaaagaaa aaagaatcaa caagggcggc gtgcaggtgg agactatctc cccaggcgac
     3061 gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg gatgcttgaa
     3121 gatggaaaga aatttgattc ctcccgggac agaaacaagc cctttaagtt tatgctaggc
     3181 aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt gggtcagaga
     3241 gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc aggcatcatc
     3301 ccaccacatg ccactctcgt ctttgatgtg gagcttctaa aactggaata acatatgaaa
     3361 taaaatacga aatgcgtacg taatgatggg cgctaaatta atgatgggcg gagttgtaat
     3421 gatgggcgct cgtttaatga tgggcgtcta gctaatgatg ggcgggcgat taatgatggg
     3481 cgaaggacta atgatgggcg cagcggtaat gatgggcgat cagctaatga tgggcgacgc
     3541 gaattaattc aacattttga cacccccata atatttttcc agaattaaca gtataaattg
     3601 catctcttgt tcaagagttc cctatcactc tctttaatca ctactcacag taacctcaac
     3661 tcctgcgccg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3721 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3781 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3841 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt cgcccgctac
     3901 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3961 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4021 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4081 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4141 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4201 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4261 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     4321 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4381 gagctgtaca agtaagccta ataaagagct cagatgcatg catcagagtg tgttggtttt
     4441 ttgtgtgtcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4501 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4561 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4621 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4681 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4741 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4801 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4861 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4921 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4981 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     5041 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5101 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5161 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5221 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccggga gctgcatgtg
     5281 tcagaggttt tcaccgtcat caccgaaacg cgcaaagggc ctcgtgatac gcctattttt
     5341 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5461 gagacaataa ccctgataaa tgcttcaata a
//
