LOCUS       sequence-371449-        7120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapamycin inducible AAV vector expressing HGF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7120)
  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 7120)
  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: pMV2 AAV-RapaInducible-NFZ-HGF
FEATURES             Location/Qualifiers
     source          1..7120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(161..180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      974..1562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1463..1482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1624..1753
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1902..1928
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2658..2711
                     /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             2712..2738
                     /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(2745..2918,2919..2924,2925..3107)
                     /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             3114..3437
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3114..3434
                     /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        3639..3752
                     /label=minimal IL-2 promoter
                     /label=IL-2 promoter
                     /note="minimal promoter from the human interleukin-2 gene"
     CDS             5698..5709
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   6018..6158
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6018..6147
                     /label=AAV2 ITR
     rep_origin      6233..6688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6320..6339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6530..6551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6705..6724)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6818..6840
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6872..6890)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6958..7062
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7063..7120,1..11,12..803)
                     /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"
ORIGIN
        1 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
       61 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      121 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      181 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      241 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      301 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      361 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      421 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      481 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      541 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      601 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
      661 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
      721 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
      781 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
      841 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      901 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      961 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1021 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1081 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1141 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1201 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1261 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1321 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1381 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1441 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1501 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1561 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1621 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     1681 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     1741 actaggggtt cctgcggccg cacgcgttaa tgatgggcgc tacagtaatg atgggcggaa
     1801 ttcctcgagg tacttatata agggggtggg ggcgcgttcg tcctcagtcg cgatcgaaca
     1861 ctcgagccga gcagacgtgc ctaaccggtg ccgccaccat gcctgctgcc aagagggtca
     1921 agttggactg gcatgaaggc ctggaagagg catctcgttt gtactttggg gaaaggaacg
     1981 tgaaaggcat gtttgaggtg ctggagccct tgcatgctat gatggaacgg ggcccccaga
     2041 ctctgaagga aacatctttt aatcaggcct atggtcgtga tttaatggag gcccaagagt
     2101 ggtgcaggaa gtacatgaaa tcagggaatg tcaaggacct cacccaagcc tgggacctct
     2161 attatcatgt gttccgacga atctcaaagc agtctggatc cggatcaggc tccgagggcc
     2221 agagcgacga gagagccctg ctggaccagc tgcacaccct gctgagcaac accgacgcca
     2281 ccggcctgga ggagatcgac agagccctgg gcatccccga gctggtgaac cagggccagg
     2341 ccctggagcc caagcaggga ggaggaggta gcggacacga gaagttcccc agcgacctgg
     2401 acctggacat gttcaacggc agcctggagt gcgacatgga gagcatcatc agaagcgagc
     2461 tgatggacgc cgacggcctg gacttcaact tcgacagcgg aggaggaggt agcggattcg
     2521 tgaccctgaa ggacgtgggc atggacttca ccctgggcga ctgggagcag ctgggcctgg
     2581 agcagggcga caccttctgg gacaccgccc tggacaactg ccaggacctg ttcctgctgg
     2641 acccccccgg atctggagag ggaaggggaa gcctcctaac ttgcggagat gtcgaggaga
     2701 atcctggccc acctgccgct aagcgtgtca agttggactc tagagaacgc ccgtatgctt
     2761 gccctgtcga gtcctgcgat cgccgctttt ctcgctcgga tgagcttacc cgccatatcc
     2821 gcatccacac aggccagaag cccttccagt gtcgaatctg catgcgtaac ttcagtcgta
     2881 gtgaccacct taccacccac atccgcaccc acacaggcgg cggccgcagg aggaagaaac
     2941 gcaccagcat agagactaac atccgtgtgg ccttagagaa gagtttcttg gagaatcaaa
     3001 agcctacctc ggaagagatc actatgattg ctgatcagct caatatggaa aaagaggtga
     3061 ttcgtgtttg gttctgtaac cgccgccaga aagaaaaaag aatcaacaag ggcggcgtgc
     3121 aggtggagac tatctcccca ggcgacgggc gcaccttccc caagcgcggc cagacctgcg
     3181 tggtgcacta caccgggatg cttgaagatg gaaagaaatt tgattcctcc cgggacagaa
     3241 acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg
     3301 ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg
     3361 gtgccactgg gcacccaggc atcatcccac cacatgccac tctcgtcttt gatgtggagc
     3421 ttctaaaact ggaataacat atgaaataaa atacgaaatg cgtacgtaat gatgggcgct
     3481 aaattaatga tgggcggagt tgtaatgatg ggcgctcgtt taatgatggg cgtctagcta
     3541 atgatgggcg ggcgattaat gatgggcgaa ggactaatga tgggcgcagc ggtaatgatg
     3601 ggcgatcagc taatgatggg cgacgcgaat taattcaaca ttttgacacc cccataatat
     3661 ttttccagaa ttaacagtat aaattgcatc tcttgttcaa gagttcccta tcactctctt
     3721 taatcactac tcacagtaac ctcaactcct gcgccgccac catgtgggtg accaaactcc
     3781 tgccagccct gctgctgcag catgtcctcc tgcatctcct cctgctcccc atcgccatcc
     3841 cctatgcaga gggacaaagg aaaagaagaa atacaattca tgaattcaaa aaatcagcaa
     3901 agactaccct aatcaaaata gatccagcac tgaagataaa aaccaaaaaa gtgaatactg
     3961 cagaccaatg tgctaataga tgtactagga ataaaggact tccattcact tgcaaggctt
     4021 ttgtttttga taaagcaaga aaacaatgcc tctggttccc cttcaatagc atgtcaagtg
     4081 gagtgaaaaa agaatttggc catgaatttg acctctatga aaacaaagac tacattagaa
     4141 actgcatcat tggtaaagga cgcagctaca agggaacagt atctatcact aagagtggca
     4201 tcaaatgtca gccctggagt tccatgatac cacacgaaca cagctttttg ccttcgagct
     4261 atcggggtaa agacctacag gaaaactact gtcgaaatcc tcgaggggaa gaagggggac
     4321 cctggtgttt cacaagcaat ccagaggtac gctacgaagt ctgtgacatt cctcagtgtt
     4381 cagaagttga atgcatgacc tgcaatgggg agagttatcg aggtctcatg gatcatacag
     4441 aatcaggcaa gatttgtcag cgctgggatc atcagacacc acaccggcac aaattcttgc
     4501 ctgaaagata tcccgacaag ggctttgatg ataattattg ccgcaatccc gatggccagc
     4561 cgaggccatg gtgctatact cttgaccctc acacccgctg ggagtactgt gcaattaaaa
     4621 catgcgctga caatactatg aatgacactg atgttccttt ggaaacaact gaatgcatcc
     4681 aaggtcaagg agaaggctac aggggcactg tcaataccat ttggaatgga attccatgtc
     4741 agcgttggga ttctcagtat cctcacgagc atgacatgac tcctgaaaat ttcaagtgca
     4801 aggacctacg agaaaattac tgccgaaatc cagatgggtc tgaatcaccc tggtgtttta
     4861 ccactgatcc aaacatccga gttggctact gctcccaaat tccaaactgt gatatgtcac
     4921 atggacaaga ttgttatcgt gggaatggca aaaattatat gggcaactta tcccaaacaa
     4981 gatctggact aacatgttca atgtgggaca agaacatgga agacttacat cgtcatatct
     5041 tctgggaacc agatgcaagt aagctgaatg agaattactg ccgaaatcca gatgatgatg
     5101 ctcatggacc ctggtgctac acgggaaatc cactcattcc ttgggattat tgccctattt
     5161 ctcgttgtga aggtgatacc acacctacaa tagtcaattt agaccatccc gtaatatctt
     5221 gtgccaaaac gaaacaattg cgagttgtaa atgggattcc aacacgaaca aacataggat
     5281 ggatggttag tttgagatac agaaataaac atatctgcgg aggatcattg ataaaggaga
     5341 gttgggttct tactgcacga cagtgtttcc cttctcgaga cttgaaagat tatgaagctt
     5401 ggcttggaat tcatgatgtc cacggaagag gagatgagaa atgcaaacag gttctcaatg
     5461 tttcccagct ggtatatggc cctgaaggat cagatctggt tttaatgaag cttgccaggc
     5521 ctgctgtcct ggatgatttt gttagtacga ttgatttacc taattatgga tgcacaattc
     5581 ctgaaaagac cagttgcagt gtttatggct ggggctacac tggattgatc aactatgatg
     5641 gcctattacg agtggcacat ctctatataa tgggaaatga gaaatgcagc cagcatcatc
     5701 gagggaaggt gactctgaat gagtctgaaa tatgtgctgg ggctgaaaag attggatcag
     5761 gaccatgtga gggggattat ggtggcccac ttgtttgtga gcaacataaa atgagaatgg
     5821 ttcttggtgt cattgttcct ggtcgtggat gtgccattcc aaatcgtcct ggtatttttg
     5881 tccgagtagc atattatgca aaatggatac acaaaattat tttaacatat aaggtaccac
     5941 agtcataggc ctaataaaga gctcagatgc atgcatcaga gtgtgttggt tttttgtgtg
     6001 tcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     6061 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     6121 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     6181 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     6241 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6301 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6361 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6421 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     6481 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6541 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     6601 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6661 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     6721 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     6781 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg ggagctgcat gtgtcagagg
     6841 ttttcaccgt catcaccgaa acgcgcaaag ggcctcgtga tacgcctatt tttataggtt
     6901 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6961 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     7021 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     7081 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
//
