LOCUS       sequence-418042-        6122 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV vector expressing tetracycline-controlled transactivator (tTA) 
            under CamK2 promoter (for expressing tTA in glutamatergic neurons).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  AUTHORS   Thompson A, Arano R, Saleem U, Preciado R, Munoz L, Nelson I, Ramos 
            K, Kim Y, Li Y, Xu W
  TITLE     Brain-wide circuit-specific targeting of astrocytes.
  JOURNAL   Cell Rep Methods. 2023 Dec 18;3(12):100653. doi: 
            10.1016/j.crmeth.2023.100653. Epub 2023 Dec 4.
  PUBMED    38052209
REFERENCE   2  (bases 1 to 6122)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep 
            Methods."; date: "2023-12-18"; volume: "3(12):100653. doi"; pages: "
            10.1016/j.crmeth.2023.100653. Epub 2023 Dec 4"
COMMENT     Sequence Label: AAV-CamK2-tTA
FEATURES             Location/Qualifiers
     source          1..6122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   670..799
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        818..2106
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             join(2147..2764,2765..2770,2771..2806,2807..2809,2810..2845,
                     2846..2848,2849..2884,2885..2893)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    2918..3506
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2971..2991)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3389..3400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3538..4014
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3737..3756)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4054..4194
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4054..4183
                     /label=AAV2 ITR
     rep_origin      4269..4724
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4356..4375)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4566..4587
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4741..4760)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4860..4882
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4920..4938)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5006..5110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5111..5179,5180..5971)
                     /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(5329..5348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg
      721 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
      781 tccatcacta ggggttcctg cggccgcacg cgtttaacat tatggcctta ggtcacttca
      841 tctccatggg gttcttcttc tgattttcta gaaaatgaga tgggggtgca gagagcttcc
      901 tcagtgacct gcccagggtc acatcagaaa tgtcagagct agaacttgaa ctcagattac
      961 taatcttaaa ttccatgcct tgggggcatg caagtacgat atacagaagg agtgaactca
     1021 ttagggcaga tgaccaatga gtttaggaaa gaagagtcca gggcagggta catctacacc
     1081 acccgcccag ccctgggtga gtccagccac gttcacctca ttatagttgc ctctctccag
     1141 tcctaccttg acgggaagca caagcagaaa ctgggacagg agccccagga gaccaaatct
     1201 tcatggtccc tctgggagga tgggtgggga gagctgtggc agaggcctca ggaggggccc
     1261 tgctgctcag tggtgacaga taggggtgag aaagcagaca gagtcattcc gtcagcattc
     1321 tgggtctgtt tggtacttct tctcacgcta aggtggcggt gtgatatgca caatggctaa
     1381 aaagcaggga gagctggaaa gaaacaagga cagagacaga ggccaagtca accagaccaa
     1441 ttcccagagg aagcaaagaa accattacag agactacaag ggggaaggga aggagagatg
     1501 aattagcttc ccctgtaaac cttagaaccc agctgttgcc agggcaacgg ggcaatacct
     1561 gtctcttcag aggagatgaa gttgccaggg taactacatc ctgtctttct caaggaccat
     1621 cccagaatgt ggcacccact agccgttacc atagcaactg cctctttgcc ccacttaatc
     1681 ccatcccgtc tgttaaaagg gccctatagt tggaggtggg ggaggtagga agagcgatga
     1741 tcacttgtgg actaagtttg ttcgcatccc cttctccaac cccctcagta catcaccctg
     1801 ggggaacagg gtccacttgc tcctgggccc acacagtcct gcagtattgt gtatataagg
     1861 ccagggcaaa gaggagcagg ttttaaagtg aaaggcaggc aggtgttggg gaggcagtta
     1921 ccggggcaac gggaacaggg cgtttcggag gtggttgcca tggggacctg gatgctgacg
     1981 aaggctcgcg aggctgtgag cagccacagt gccctgctca gaagccccaa gctcgtcagt
     2041 caagccggtt ctccgtttgc actcaggagc acgggcaggc gagtggcccc tagttctggg
     2101 ggcagctcta gagcggtacc ggatccccgg gtaccggtag gccaccatgt ctagactgga
     2161 caagagcaaa gtcataaact ctgctctgga attactcaat gaagtcggta tcgaaggcct
     2221 gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt
     2281 gaagaacaag cgggccctgc tcgatgccct ggcaatcgag atgctggaca ggcatcatac
     2341 ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc
     2401 attccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc
     2461 aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt
     2521 ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt
     2581 attggaggat caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc
     2641 tatgccccca cttctgagac aagcaattga gctgttcgac catcagggag ccgaacctgc
     2701 cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag
     2761 cggcgggccg gccgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct
     2821 tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat
     2881 gctccccggg taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     2941 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3001 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3061 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3121 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggaattg ccaccacctg
     3181 tcagctcctt tccggaactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3241 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3301 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3361 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3421 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3481 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
     3541 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3601 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3661 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3721 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3781 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     3841 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3901 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3961 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     4021 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     4081 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4141 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4201 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4261 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4321 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4381 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4441 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4501 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4561 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4621 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4681 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4741 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4801 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4861 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4921 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4981 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5041 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5101 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5161 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5221 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5281 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5341 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5401 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5461 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5521 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5581 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5641 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5701 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5761 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5821 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5881 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5941 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6001 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6061 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6121 ag
//
