LOCUS       sequence-429499-        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of Cre- and doxycycline-dependent
            expression of HA-epitope tagged CD4 receptor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  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 6879)
  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-CD4
FEATURES             Location/Qualifiers
     source          1..6879
                     /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             join(3732..3806,3807..5105)
                     /codon_start=1
                     /product="human CD4 cell surface glycoprotein"
                     /label=CD4
                     /translation="MNRGVPFRHLLLVLQLALLPAATQGKKVVLGKKGDTVELTCTASQ
                     KKSIQFHWKNSNQIKILGNQGSFLTKGPSKLNDRADSRRSLWDQGNFPLIIKNLKIEDS
                     DTYICEVEDQKEEVQLLVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRGKNI
                     QGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFQKASSIVYKKEGEQVEFS
                     FPLAFTVEKLTGSGELWWQAERASSSKSWITFDLKNKEVSVKRVTQDPKLQMGKKLPLH
                     LTLPQALPQYAGSGNLTLALEAKTGKLHQEVNLVVMRATQLQKNLTCEVWGPTSPKLML
                     SLKLENKEAKVSKREKAVWVLNPEAGMWQCLLSDSGQVLLESNIKVLPTWSTPVQPMAL
                     IVLGGVAGLLLFIGLGIFFCVRCRHRRRQAERMSQIKRLLSEKKTCQCPHRFQKTCSPI
                     "
     CDS             5115..5141
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5121..5147
                     /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    5179..5640
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(6143..6176)
                     /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(6206..6230)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6227..6265)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    6277..6295
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6313..6331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6348..6366
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6384..6402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6420..6438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6455..6473
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(6512..6531)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    6585..6633
                     /note="synthetic polyadenylation signal"
     repeat_region   6689..6818
                     /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 taactagtaa catgaaccgg ggagtccctt ttaggcactt gcttctggtg ctgcaactgg
     3781 cgctcctccc agcagccact cagggaaaga aagtggtgct gggcaaaaaa ggggatacag
     3841 tggaactgac ctgtacagct tcccagaaga agagcataca attccactgg aaaaactcca
     3901 accagataaa gattctggga aatcagggct ccttcttaac taaaggtcca tccaagctga
     3961 atgatcgcgc tgactcaaga agaagccttt gggaccaagg aaactttccc ctgatcatca
     4021 agaatcttaa gatagaagac tcagatactt acatctgtga agtggaggac cagaaggagg
     4081 aggtgcaatt gctagtgttc ggattgactg ccaactctga cacccacctg cttcaggggc
     4141 agagcctgac cctgaccttg gagagccccc ctggtagtag cccctcagtg caatgtagga
     4201 gtccaagggg taaaaacata caggggggga agaccctctc cgtgtctcag ctggagctcc
     4261 aggatagtgg cacctggaca tgcactgtct tgcagaacca gaagaaggtg gagttcaaaa
     4321 tagacatcgt ggtgctagct ttccagaagg cctccagcat agtctataag aaagaggggg
     4381 aacaggtgga gttctccttc ccactcgcct ttacagttga aaagctgacg ggcagtggcg
     4441 agctgtggtg gcaggcggag agggcttcct cctccaagtc ttggatcacc tttgacctga
     4501 agaacaagga agtgtctgta aaacgggtta cccaggaccc taagctccag atgggcaaga
     4561 agctcccgct ccacctcacc ctgccccagg ccttgcctca gtatgctggc tctggaaacc
     4621 tcaccctggc ccttgaagcg aaaacaggaa agttgcatca ggaagtgaac ctggtggtga
     4681 tgagagccac tcagctccag aaaaatttga cctgtgaggt gtggggaccc acctccccta
     4741 agctgatgct gagcttgaaa ctggagaaca aggaggcaaa ggtctcgaag cgggagaagg
     4801 cggtgtgggt gctgaaccct gaggcgggga tgtggcagtg tctgctgagt gactcgggac
     4861 aggtcctgct ggaatccaac atcaaggttc tgcccacatg gtccaccccg gtgcagccaa
     4921 tggccctgat tgtgctgggg ggcgtcgccg gcctcctgct tttcattggg ctaggcatct
     4981 tcttctgtgt caggtgccgg caccgaaggc gccaagcaga gcggatgtct cagatcaaga
     5041 gactcctcag tgagaagaag acctgccagt gccctcaccg gtttcagaag acatgtagcc
     5101 ccattctcga gggttatccg tatgatgttc ctgattatgc tagcctctag gtcgacggcg
     5161 cgcctataaa acgatccgct gtaagtctgc agaaattgat gatctattaa acaataaaga
     5221 tgtccactaa aatggaagtt tttcctgtca tactttgtta agaagggtga gaacagagta
     5281 cctacatttt gaatggaagg attggagcta cgggggtggg ggtggggtgg gattagataa
     5341 atgcctgctc tttactgaag gctctttact attgctttat gataatgttt catagttgga
     5401 tatcataatt taaacaagca aaaccaaatt aagggccagc tcattcctcc cactcatgat
     5461 ctatagatct atagatctct cgtgggatca ttgtttttct cttgattccc actttgtggt
     5521 tctaagtact gtggtttcca aatgtgtcag tttcatagcc tgaagaacga gatcagcagc
     5581 ctctgttcca catacacttc attctcagta ttgttttgcc aagttctaat tccatcagaa
     5641 agcttgcaga agatctcccc aactgggtcg agcaagtgag tgcaagtggg ttttaggacc
     5701 aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac
     5761 ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg
     5821 cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct
     5881 ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc
     5941 ccgcaaactc cccttcccgg ccaccttggt cgcgcccgtg cccccctatc tcgcgcctcg
     6001 cgtggtgcgg tccggctggg ccggcggcgg cgactcagcg ctgcctcagt ctgcggtggg
     6061 cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat tcgatcaatt gcatttccta
     6121 gcggatacaa caattgcatt tctaccgttc gtatagcata cattatacga agttatcatg
     6181 cgtcgtacac tgaaatcgaa ttcgcgatct gacggttcac taaacgagct ctgcttatat
     6241 aggcctccca ccgtacacgc ctacctcgac atacgttctc tatcactgat agggagtaaa
     6301 ctcgacatac gttctctatc actgataggg ataaactcga catacgttct ctatcactga
     6361 tagggagtaa actcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt
     6421 ctctatcact gatagggagt aaactcgaca tcgttctcta tcactgatag ggagtaaact
     6481 cgggtaccta gagcatgggg cccttgcact ggggacagcc tattttgcta gtttgttttg
     6541 tttcgttttg ttttgatgga gagcgtatgt tagtactatc gattcacaca aaaaaccaac
     6601 acacagatgt aatgaaaata aagatatttt attgcggccg ctccaagtac ctcccgtacc
     6661 ttaatatttc tagaacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc
     6721 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     6781 ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca
     6841 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgtt
//
