LOCUS       sequence-343534-        5598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of InteinC-rtTAC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5598)
  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 5598)
  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-rtTAc
FEATURES             Location/Qualifiers
     source          1..5598
                     /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"
     CDS             1587..1697
                     /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"
     misc_feature    1725..2313
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1778..1798)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2196..2207)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2345..2821
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2544..2563)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2861..3001
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2861..2990
                     /label=AAV2 ITR
     rep_origin      3076..3531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3163..3182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3373..3394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3548..3567)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3667..3689
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3727..3745)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3813..3917
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3918..3986,3987..4778)
                     /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(4136..4155)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4949..5537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5438..5457
                     /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 ctctagagga tccccgggta
     1441 ccggtgccac catgatgctg aagaagatcc tgaagatcga ggagctggac gagcgcgagc
     1501 tgatcgacat cgaggtgagc ggcaaccacc tgttctacgc caacgacatc ctgacccaca
     1561 acagcagcag cagcgacgtg ccgaccgacg cccttgacga ttttgactta gacatgctcc
     1621 cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt
     1681 ttgaccttga catgctcccc gggtaagata tcaagcttat cgataatcaa cctctggatt
     1741 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     1801 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     1861 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     1921 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     1981 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2041 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2101 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     2161 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2221 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     2281 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     2341 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     2401 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     2461 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     2521 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     2581 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     2641 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     2701 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     2761 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     2821 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     2881 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2941 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3001 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3061 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3121 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     3181 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3241 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3301 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3361 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     3421 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     3481 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     3541 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3601 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3661 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3721 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3781 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3841 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3901 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3961 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4021 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4081 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4141 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4201 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4261 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4321 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4381 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4441 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4501 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4561 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4621 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4681 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4741 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4801 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4861 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4921 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4981 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5041 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5101 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5161 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5221 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5281 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5341 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5401 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5461 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5521 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5581 ggccttttgc tcacatgt
//
