LOCUS       sequence-72981-e        5810 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV TALE synthesis backbone with NI half repeat; fused to nuclear 
            localization sequence linker and SID4X transcriptional repressor 
            effector domain. Synapsin promoter for neuronal expression. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  AUTHORS   Konermann S, Brigham MD, Trevino A, Hsu PD, Heidenreich M, Le Cong, 
            Platt RJ, Scott DA, Church GM, Zhang F
  TITLE     Optical control of mammalian endogenous transcription and epigenetic
            states.
  JOURNAL   Nature. 2013 Jul 23. doi: 10.1038/nature12466.
  PUBMED    23877069
REFERENCE   2  (bases 1 to 5810)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2013 Jul 23. doi: 10.1038/nature12466."
COMMENT     Sequence Label: 
            pAAV_hSyn_TALEBB(NI)-NLS-SID4X_2A_phiLOV2.1_WPRE_bGHpA
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             651..677
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1370..1390
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1790..1843
                     /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"
     misc_feature    2201..2789
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2254..2274)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2672..2683)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2831..2848)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2837..3044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3073..3213
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3073..3202
                     /label=AAV2 ITR
     rep_origin      3288..3743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3375..3394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3585..3606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3760..3779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3879..3901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3939..3957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4025..4129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4130..4198,4199..4990)
                     /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(4348..4367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5161..5749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5650..5669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg tacccatacg
      661 atgttccaga ttacgctgta gatttgagaa ctttgggata ttcacagcag cagcaggaaa
      721 agatcaagcc caaagtgagg tcgacagtcg cgcagcatca cgaagcgctg gtgggtcatg
      781 ggtttacaca tgcccacatc gtagccttgt cgcagcaccc tgcagccctt ggcacggtcg
      841 ccgtcaagta ccaggacatg attgcggcgt tgccggaagc cacacatgag gcgatcgtcg
      901 gtgtggggaa acagtggagc ggagcccgag cgcttgaggc cctgttgacg gtcgcgggag
      961 agctgagagg gcctcccctt cagctggaca cgggccagtt gctgaagatc gcgaagcggg
     1021 gaggagtcac ggcggtcgag gcggtgcacg cgtggcgcaa tgcgctcacg ggagcacccc
     1081 tcaacctgac agagacccgt acgggtctcg actcacgcct gagcaggtag tggctattgc
     1141 atccaatatc gggggcagac ccgcactgga gtcaatcgtg gcccagcttt cgaggccgga
     1201 ccccgcgctg gccgcactca ctaatgatca tcttgtagcg ctggcctgcc tcggcggacg
     1261 acccgccttg gatgcggtga agaaggggct cccgcacgcg cctgcattga ttaagcggac
     1321 caacagaagg attcccgaga ggacatcaca tcgagtggca gctagcagcc ccaagaagaa
     1381 gagaaaggtg gaggccagca tgaacatcca gatgctgctg gaggccgctg actacctgga
     1441 acggagagag cgcgaagccg agcacggata tgcttcaatg ctgcccggaa gcggcatgaa
     1501 tattcagatg ctgctggagg ctgctgatta cctggaaagg cgcgaacggg aggccgaaca
     1561 tggctatgct tccatgctgc ctgggtctgg aatgaatatc caaatgctgc tggaggcagc
     1621 cgattacctg gaacggagag aaagagaagc cgagcacgga tacgccagca tgctgccagg
     1681 cagcgggatg aacatacaaa tgctgctgga ggctgccgat tacctggaga ggcgcgagag
     1741 agaagctgaa catggctatg cctctatgct gcccccatgg ggcagtggag agggcagagg
     1801 aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaatcgaga agtccttcgt
     1861 aataacagat ccaagactgc ctgattaccc catcattttt gccagcgacg gttttctgga
     1921 gctcaccgaa tattccaggg aggaaatcat ggggagaaac gccaggttcc tgcaagggcc
     1981 tgagacagac caggccaccg ttcagaagat tcgcgatgct atccgcgatc agagggaaac
     2041 cactgtgcag ttaataaact acactaagtc aggcaaaaag ttctggaatc tgcttcatct
     2101 ccaacctgtc cgggatcgaa agggagggct gcaatacttc atcggtgtgc agctggtggg
     2161 aagtgatcat gtgtaagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     2221 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2281 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2341 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2401 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2461 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2521 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2581 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     2641 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2701 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     2761 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agctagagct
     2821 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2881 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2941 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3001 agcaaggggg aggattggga agacaatagc aggcatgctg gggaggtaac cacgtgcgga
     3061 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3121 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3181 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3241 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3421 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3481 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3601 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     3661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3721 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3781 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3841 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3901 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3961 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4021 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4081 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4141 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4201 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4261 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4321 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4381 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4441 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4501 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4561 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4621 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4681 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4741 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4801 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     4861 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4921 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4981 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5041 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5101 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5161 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5221 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5281 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5341 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5401 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5461 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5521 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5581 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5641 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5701 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5761 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
