LOCUS       sequence-376508-        7291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hM3D(Gq) under the hSyn promoter, and d2tTA under the pRAM
            (synthetic Fos) promoter. Plasmid 1 for recording Fos expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7291)
  AUTHORS   Lee S, Nouraein S, Kwon JJ, Huang Z, Wojick JA, Xia B, Corder G, 
            Szablowski JO
  TITLE     Engineered serum markers for non-invasive monitoring of gene 
            expression in the brain.
  JOURNAL   Nat Biotechnol. 2024 Jan 10. doi: 10.1038/s41587-023-02087-x.
  PUBMED    38200117
REFERENCE   2  (bases 1 to 7291)
  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 
            Biotechnol. 2024 Jan 10. doi: 10.1038/s41587-023-02087-x."
COMMENT     Sequence Label: pAAV-hSyn-hM3D(Gq)::RAM-d2tTA
FEATURES             Location/Qualifiers
     source          1..7291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     472..491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   633..762
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        807..1254
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1295..1321
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3164..3184)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(3394..3411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3400..3511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3659..3707
                     /note="synthetic polyadenylation signal"
     misc_feature    3721..3812
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3761..3780
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             join(4178..4795,4796..4801,4802..4837,4838..4840,4841..4876,
                     4877..4879,4880..4915,4916..4924)
                     /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"
     primer_bind     complement(4937..4953)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4978..4997)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4979..4997)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5023..5157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5060..5079)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5114..5133
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5186..5326
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5186..5315
                     /label=AAV2 ITR
     rep_origin      5401..5856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5488..5507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5698..5719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5873..5892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5992..6014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6138..6242
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6243..6311,6312..7103)
                     /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(6461..6480)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7274..7291,1..571)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
       61 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
      121 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
      181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
      241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
      421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      601 cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct
      661 cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
      721 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtgtgt
      781 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
      841 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
      901 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
      961 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
     1021 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
     1081 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
     1141 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
     1201 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga
     1261 aggtaccgga tcctctagag tcgacgccac catgtaccca tacgatgttc cagattacgc
     1321 taccttgcac aataacagta caacctcgcc tttgtttcca aacatcagct cctcctggat
     1381 acacagcccc tccgatgcag ggctgccccc gggaaccgtc actcatttcg gcagctacaa
     1441 tgtttctcga gcagctggca atttctcctc tccagacggt accaccgatg accctctggg
     1501 aggtcatacc gtctggcaag tggtcttcat cgctttctta acgggcatcc tggccttggt
     1561 gaccatcatc ggcaacatcc tggtaattgt gtcatttaag gtcaacaagc agctgaagac
     1621 ggtcaacaac tacttcctct taagcctggc ctgtgccgat ctgattatcg gggtcatttc
     1681 aatgaatctg tttacgacct acatcatcat gaatcgatgg gccttaggga acttggcctg
     1741 tgacctctgg cttgccattg actgcgtagc cagcaatgcc tctgttatga atcttctggt
     1801 catcagcttt gacagatact tttccatcac gaggccgctc acgtaccgag ccaaacgaac
     1861 aacaaagaga gccggtgtga tgatcggtct ggcttgggtc atctcctttg tcctttgggc
     1921 tcctgccatc ttgttctggc aatactttgt tggaaagaga actgtgcctc cgggagagtg
     1981 cttcattcag ttcctcagtg agcccaccat tacttttggc acagccatcg ctggttttta
     2041 tatgcctgtc accattatga ctattttata ctggaggatc tataaggaaa ctgaaaagcg
     2101 taccaaagag cttgctggcc tgcaagcctc tgggacagag gcagagacag aaaactttgt
     2161 ccaccccacg ggcagttctc gaagctgcag cagttacgaa cttcaacagc aaagcatgaa
     2221 acgctccaac aggaggaagt atggccgctg ccacttctgg ttcacaacca agagctggaa
     2281 acccagctcc gagcagatgg accaagacca cagcagcagt gacagttgga acaacaatga
     2341 tgctgctgcc tccctggaga actccgcctc ctccgacgag gaggacattg gctccgagac
     2401 gagagccatc tactccatcg tgctcaagct tccgggtcac agcaccatcc tcaactccac
     2461 caagttaccc tcatcggaca acctgcaggt gcctgaggag gagctgggga tggtggactt
     2521 ggagaggaaa gccgacaagc tgcaggccca gaagagcgtg gacgatggag gcagttttcc
     2581 aaaaagcttc tccaagcttc ccatccagct agagtcagcc gtggacacag ctaagacttc
     2641 tgacgtcaac tcctcagtgg gtaagagcac ggccactcta cctctgtcct tcaaggaagc
     2701 cactctggcc aagaggtttg ctctgaagac cagaagtcag atcactaagc ggaaaaggat
     2761 gtccctggtc aaggagaaga aagcggccca gaccctcagt gcgatcttgc ttgccttcat
     2821 catcacttgg accccataca acatcatggt tctggtgaac accttttgtg acagctgcat
     2881 acccaaaacc ttttggaatc tgggctactg gctgtgctac atcaacagca ccgtgaaccc
     2941 cgtgtgctat gctctgtgca acaaaacatt cagaaccact ttcaagatgc tgctgctgtg
     3001 ccagtgtgac aaaaaaaaga ggcgcaagca gcagtaccag cagagacagt cggtcatttt
     3061 tcacaagcgc gcacccgagc aggccttgta aatcgaattc cgctcgagat aatcaacctc
     3121 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3181 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3241 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     3301 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttct
     3361 gatcgatacc gtcgactaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     3421 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3481 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3541 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcagactag
     3601 tacgtctctc aaggataagt aagtaatatt aaggtacggg aggtattgga caggccgcaa
     3661 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact
     3721 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
     3781 cagtgcaagt gcaggtgcca gaacatttct ctggcctaac tggccgacgc gtaccctaga
     3841 agtttgttcg tgactcagac tagaagtttg ttcgtgactc agactagaag tttgttcgtg
     3901 actcagacta gaagtttgtt cgtgactcag aactcattca taaaacgctt gttataaaag
     3961 cagtggctgc ggcgcctcgt actccaaccg catctgcagc gagcaactga gaagccaaga
     4021 ctgagccggc ggccggatcc agctcatgag ccatggcttc ccgccggagg tggaggagca
     4081 ggatgatggc acgctgccca tgtcttgtgc ccaggagagc gggatggacc gtcaccctgc
     4141 agcctgtgct tctgctagga tcaatgtgga attcaccatg tctagactgg acaagagcaa
     4201 agtcataaac tctgctctgg aattactcaa tgaagtcggt atcgaaggcc tgacgacaag
     4261 gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa
     4321 gcgggccctg ctcgatgccc tggcaatcga gatgctggac aggcatcata cccacttctg
     4381 ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cattccgctg
     4441 tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa
     4501 acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga
     4561 gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga
     4621 tcaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc
     4681 acttctgaga caagcaattg agctgttcga ccatcaggga gccgaacctg ccttcctttt
     4741 cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc
     4801 ggccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt
     4861 tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg
     4921 gtaactaagt aagcatcgat accgtcgacc tcgagggggg gcccggtacc caattcgccc
     4981 tatagtgagt cgtattacgc gcgcagcggc cgaccatggc ccaacttgtt tattgcagct
     5041 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5101 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctcc
     5161 ggacacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     5221 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     5281 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     5341 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     5401 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5461 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5521 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5581 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     5641 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5701 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     5761 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     5821 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     5881 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5941 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     6001 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     6061 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     6121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6181 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6241 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6301 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6361 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6421 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6481 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6541 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6601 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6661 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6721 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6781 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6841 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6901 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6961 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7021 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7081 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7141 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7201 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7261 tcaaaggatc ttcttgagat cctttttttc t
//
