LOCUS       sequence-343532-        5848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of InteinC-tTAC in Cre recombinase positive cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5848)
  AUTHORS   Chen HS, Zhang XL, Yang RR, Wang GL, Zhu XY, Xu YF, Wang DY, Zhang 
            N, Qiu S, Zhan LJ, Shen ZM, Xu XH, Long G, Xu C
  TITLE     An intein-split transactivator for intersectional neural imaging and
            optogenetic manipulation.
  JOURNAL   Nat Commun. 2022 Jun 23;13(1):3605. doi: 10.1038/s41467-022-31255-x.
  PUBMED    35739125
REFERENCE   2  (bases 1 to 5848)
  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 
            Commun."; date: "2022-06-23"; volume: "13(1):3605. doi"; pages: " 
            10.1038/s41467-022-31255-x"
COMMENT     Sequence Label: pAAV-pEF1a-DIO-tTAc
FEATURES             Location/Qualifiers
     source          1..5848
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1575..1685)
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     protein_bind    1833..1866
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1917..1950
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1975..2563
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2028..2048)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2446..2457)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2595..3071
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2794..2813)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3111..3251
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3111..3240
                     /label=AAV2 ITR
     rep_origin      3326..3781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3413..3432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3623..3644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3798..3817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3917..3939
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3977..3995)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4063..4167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4168..4236,4237..5028)
                     /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(4386..4405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5199..5787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5688..5707
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttacc cggggagcat gtcaaggtca aaatcgtcaa gagcgtcagc aggcagcata
     1621 tcaaggtcaa agtcgtcaag ggcatcggct gggagcatgt ctaagtcaaa atcgtcaagg
     1681 gcgtcggccg gcacgtcgct gctgctgctg ttgtgggtca ggatgtcgtt ggcgtagaac
     1741 aggtggttgc cgctcacctc gatgtcgatc agctcgcgct cgtccagctc ctcgatcttc
     1801 aggatcttct tcagcatcat ggtggcgcta gcataacttc gtataaagta tcctatacga
     1861 agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa
     1921 cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat cgataatcaa
     1981 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2041 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2101 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2161 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2221 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2281 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2341 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     2401 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2461 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2521 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     2581 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     2641 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     2701 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     2761 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     2821 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     2881 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     2941 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     3001 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     3061 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     3121 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3181 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3241 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3301 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     3421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3541 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3661 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     3721 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     3781 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3841 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3901 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3961 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4021 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4081 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4141 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4201 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4261 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4321 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4381 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4441 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4501 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4561 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4621 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4681 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4741 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4801 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4861 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4921 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4981 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5041 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5101 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5341 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5761 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5821 gccttttgct ggccttttgc tcacatgt
//
