LOCUS       sequence-429501-        7101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of Cre- and doxycycline-dependent
            expression of HA-epitope tagged constitutively-active BMPR1A 
            receptor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7101)
  AUTHORS   Okur Z, Schlauri N, Bitsikas V, Panopoulou M, Karmakar K, Schreiner 
            D, Scheiffele P
  TITLE     Control of neuronal excitation-inhibition balance by BMP-SMAD1 
            signaling
  JOURNAL   bioRxiv 2023
REFERENCE   2  (bases 1 to 7101)
  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: "bioRxiv 
            2023"
COMMENT     Sequence Label: pAAV-TetON-Bmpr1A-ca
FEATURES             Location/Qualifiers
     source          1..7101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1793..1811
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1849..1871)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1971..1990
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2007..2462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2144..2165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2356..2375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2537..2677
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2548..2677
                     /label=AAV2 ITR
     polyA_signal    2789..2900
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2889..2906
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2930..3637)
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     protein_bind    3658..3691
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             5337..5363
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5343..5369
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     polyA_signal    5401..5862
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(6365..6398)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(6428..6452)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6449..6487)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    6499..6517
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6535..6553
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6570..6588
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6606..6624
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6642..6660
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6677..6695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(6734..6753)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    6807..6855
                     /note="synthetic polyadenylation signal"
     repeat_region   6911..7040
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1801 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     1981 gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc
     2041 gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc
     2101 ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag
     2161 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2221 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     2281 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2341 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt
     2401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2461 gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2521 taccgcatca ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
     2581 gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
     2641 agagagggag tggccaactc catcactagg ggttcctgcg gccgctcggt ccgcacgtgc
     2701 atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
     2761 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
     2821 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
     2881 ctggcaacta gaaggcacag tcgaggctga tcagcgagct ctagtcgact tacccgggga
     2941 gcatgtcaag gtcaaaatcg tcaagggcat cggctgggag catgtctaag tcaaaatcgt
     3001 caagggcgtc ggtcggcccg ccgctttcgc actttagctg tttctccagg ccacatatga
     3061 ttagttccag gccgaaaagg aaggcaggtt cggctccctg ccggtcgaac agctcaattg
     3121 cttgtctcag aagtgggggc atagaatcgg tggtaggtgt ctctctttcc tcttttgcta
     3181 cttgatgctc ctgttcctcc aatacgcagc ccagtgtaaa gtggcccacg gcggacagag
     3241 cgtacagtgc gttctccagg gagaagcctt gctgacacag gaacgcgagc tgattttcca
     3301 gggtttcgta ctgtttctct gttgggcggg tgccgagatg cactttagcc ccgtcgcgat
     3361 gtgagaggag agcacagcgg tatgacttgg cgttgttccg cagaaagtct tgccatgact
     3421 cgccttccag ggggcagaag tgggtatgat gcctgtccag catctcgatt ggcagggcat
     3481 cgagcagggc ccgcttgttc ttcacgtgcc agtacagggt aggctgctca actcccagct
     3541 tttgagcgag tttccttgtc gtcaggcctt cgataccgac accattgagt aattccagag
     3601 ctccgtttat gactttgctc ttgtccagcc tagacatggt ggcaagcttg cggccgctac
     3661 cgttcgtata atgtatgcta tacgaagtta tcatagcgct agatatgagc tagcgtttta
     3721 taactagtac catggctcag ctatacattt acatcagatt attgggagcc tatttgttca
     3781 tcatttctcg tgttcaagga cagaatctgg atagtatgct tcatggcact gggatgaaat
     3841 cagactccga ccagaaaaag tcagaaaatg gagtaacctt agcaccagag gataccttgc
     3901 cttttttaaa gtgctattgc tcagggcact gtccagatga tgctattaat aacacatgca
     3961 taactaatgg acattgcttt gccatcatag aagaagatga ccagggagaa accacattag
     4021 cttcagggtg tatgaaatat gaaggatctg attttcagtg caaagattct ccaaaagccc
     4081 agctacgccg gacaatagaa tgttgtcgga ccaatttatg taaccagtat ttgcaaccca
     4141 cactgccccc tgttgtcata ggtccgtttt ttgatggcag cattcgatgg ctggttttgc
     4201 tcatttctat ggctgtctgc ataattgcta tgatcatctt ctccagctgc ttttgttaca
     4261 aacattattg caagagcatc tcaagcagac gtcgttacaa tcgtgatttg gaacaggatg
     4321 aagcatttat tccagttgga gaatcactaa aagaccttat tgaccagtca caaagttctg
     4381 gtagtgggtc tggactacct ttattggttc agcgaactat cgcgaaagat atccagatgg
     4441 tccggcaagt tggtaaaggc cgatatggag aagtatggat gggcaaatgg cgtggcgaaa
     4501 aagtggcggt gaaagtattc tttaccactg aagaagccag ctggtttcga gaaacagaaa
     4561 tctaccaaac tgtgctaatg cgccatgaaa acatacttgg tttcatagcg gcagacatta
     4621 aaggtacagg ttcctggact cagctctatt tgattactga ttaccatgaa aatggatctc
     4681 tctatgactt cctgaaatgt gctacactgg acaccagagc cctgcttaaa ttggcttatt
     4741 cagctgcctg tggtctgtgc cacctgcaca cagaaattta tggcacccaa ggaaagcccg
     4801 caattgctca tcgagaccta aagagcaaaa acatcctcat caagaaaaat gggagttgct
     4861 gcattgctga cctgggcctt gctgttaaat tcaacagtga cacaaatgaa gttgatgtgc
     4921 ccttgaatac cagggtgggc accaaacgct acatggctcc cgaagtgctg gacgaaagcc
     4981 tgaacaaaaa ccacttccag ccctacatca tggctgacat ctacagcttc ggcctaatca
     5041 tttgggagat ggctcgtcgt tgtatcacag gagggatcgt ggaagaatac caattgccat
     5101 attacaacat ggtaccgagt gatccgtcat acgaagatat gcgtgaggtt gtgtgtgtca
     5161 aacgtttgcg gccaattgtg tctaatcggt ggaacagtga tgaatgtcta cgagcagttt
     5221 tgaagctaat gtcagaatgc tgggcccaca atccagcctc cagactcaca gcattgagaa
     5281 ttaagaagac gcttgccaag atggttgaat cccaagatgt aaaaatcctc gagggttatc
     5341 cgtatgatgt tcctgattat gctagcctct aggtcgacgg cgcgcctata aaacgatccg
     5401 ctgtaagtct gcagaaattg atgatctatt aaacaataaa gatgtccact aaaatggaag
     5461 tttttcctgt catactttgt taagaagggt gagaacagag tacctacatt ttgaatggaa
     5521 ggattggagc tacgggggtg ggggtggggt gggattagat aaatgcctgc tctttactga
     5581 aggctcttta ctattgcttt atgataatgt ttcatagttg gatatcataa tttaaacaag
     5641 caaaaccaaa ttaagggcca gctcattcct cccactcatg atctatagat ctatagatct
     5701 ctcgtgggat cattgttttt ctcttgattc ccactttgtg gttctaagta ctgtggtttc
     5761 caaatgtgtc agtttcatag cctgaagaac gagatcagca gcctctgttc cacatacact
     5821 tcattctcag tattgttttg ccaagttcta attccatcag aaagcttgca gaagatctcc
     5881 ccaactgggt cgagcaagtg agtgcaagtg ggttttagga ccaggatgag gcggggtggg
     5941 ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa
     6001 attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca
     6061 ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc
     6121 cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc
     6181 ggccaccttg gtcgcgcccg tgccccccta tctcgcgcct cgcgtggtgc ggtccggctg
     6241 ggccggcggc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
     6301 gtgcctgaga gcgcagtcga attcgatcaa ttgcatttcc tagcggatac aacaattgca
     6361 tttctaccgt tcgtatagca tacattatac gaagttatca tgcgtcgtac actgaaatcg
     6421 aattcgcgat ctgacggttc actaaacgag ctctgcttat ataggcctcc caccgtacac
     6481 gcctacctcg acatacgttc tctatcactg atagggagta aactcgacat acgttctcta
     6541 tcactgatag ggataaactc gacatacgtt ctctatcact gatagggagt aaactcgaca
     6601 tacgttctct atcactgata gggagtaaac tcgacatacg ttctctatca ctgataggga
     6661 gtaaactcga catcgttctc tatcactgat agggagtaaa ctcgggtacc tagagcatgg
     6721 ggcccttgca ctggggacag cctattttgc tagtttgttt tgtttcgttt tgttttgatg
     6781 gagagcgtat gttagtacta tcgattcaca caaaaaacca acacacagat gtaatgaaaa
     6841 taaagatatt ttattgcggc cgctccaagt acctcccgta ccttaatatt tctagaacgc
     6901 gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     6961 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg
     7021 agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7081 aaaggccgcg ttgctggcgt t
//
