LOCUS       sequence-314299-        7050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of HA Allatostatin-3, neurophysin, IRES mCherry under
            the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7050)
  AUTHORS   Ngo HB, Melo MR, Layfield S, Connelly AA, Bassi JK, Xie L, Menuet C,
            McDougall SJ, Bathgate RAD, Allen AM
  TITLE     A Chemogenetic Tool that Enables Functional Neural Circuit Analysis.
  JOURNAL   Cell Rep. 2020 Sep 15;32(11):108139. doi: 
            10.1016/j.celrep.2020.108139.
  PUBMED    32937120
REFERENCE   2  (bases 1 to 7050)
  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: "Cell Rep.";
            date: "2020-09-15"; volume: "32(11):108139. doi"; pages: " 
            10.1016/j.celrep.2020.108139"
COMMENT     Sequence Label: pAM-DCA-HA-Ast-3-NP-1-IRES-mCherry-WPRE
FEATURES             Location/Qualifiers
     source          1..7050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     promoter        466..743
                     /label=chicken beta-actin promoter
     primer_bind     1022..1044
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1106..1125
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1218..1244
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1594..2143
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1738..1755)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1965..1984
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(2291..2309)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2559..2578
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2876..3464
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2929..2949)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3347..3358)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3491..3508)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3497..3721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3931..4260)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3945..4080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4245..4265
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4506..5094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4586..4605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5265..6056,6057..6125))
                     /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     5888..5907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6126..6230)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6298..6316
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6354..6376)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6476..6495
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6709..6728
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7018..7036
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7018..7035
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctaggt accatcctac tactagtacg tcgtaccatt gacgtcaata atgacgtatg
      241 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      301 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      361 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      421 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      481 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      541 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccrggsg
      601 gggsggggsg gggsgrgggg sggggsgggg sgaggcggag aggtgcggcg gcagccaatc
      661 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      721 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
      781 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      841 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      901 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
      961 ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg
     1021 gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc
     1081 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
     1141 ggcaaagaat tggatccacc atggcctgcc ccagtcttgc ttgctgcctg cttggcctac
     1201 tggctctgac ctccgcctac ccatacgatg ttccagatta cgctagcaga ccctacagct
     1261 tcggcctggg caagagggct gcgctagacc tggatatgcg caagtgtctt ccctgcggac
     1321 ccggcggcaa agggcgctgc ttcgggccga gcatctgctg cgcggacgag ctgggctgct
     1381 tcgtgggcac cgccgaggcg ctgcgctgcc aggaggagaa ctacctgccc tcgccctgcc
     1441 agtctggcca gaagccttgc ggaagcggag gccgctgcgc caccgcgggc atctgctgta
     1501 gcccggatgg ctgccgcacc gaccccgcct gcgaccctga gtctgccttc tccgagcgct
     1561 gactcgagcg ggatcaattc cgcccccccc ctaacgttac tggccgaagc cgcttggaat
     1621 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     1681 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     1741 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     1801 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     1861 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     1921 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     1981 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2041 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2101 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatggt gagcatggac
     2161 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2221 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2281 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2341 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2401 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2461 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2521 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2581 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2641 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2701 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2761 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2821 cactccaccg gcggcatgga cgagctgtac aagtaagaat tcgaagctta tcgataatca
     2881 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2941 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3001 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3061 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3121 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3181 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3241 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     3301 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3361 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3421 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgact
     3481 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3541 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3601 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3661 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3721 gcttctgagg cggaaagaac cagctggggc tcgactagag catggctacg tagataagta
     3781 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     3841 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg
     3901 cctcagtgag cgagcgagcg cgcagagctt tttgcaaaag cctaggcctc caaaaaagcc
     3961 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     4021 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     4081 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     4141 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     4201 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     4261 attccacagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4321 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4381 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4441 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4501 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4561 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4621 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4681 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4741 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4801 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4861 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4921 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4981 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5041 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5101 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5161 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5221 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5281 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5341 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5401 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5461 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5521 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5581 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5641 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5701 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5761 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5821 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5881 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5941 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6001 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6061 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6121 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6181 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6241 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6301 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6361 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6421 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     6481 tcagagcaga ttgtactgag agtgcaccat tcgacgctct cccttatgcg actcctgcat
     6541 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     6601 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     6661 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     6721 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     6781 gtagaggatc tggctagcga tgaccctgct gattggttcg ctgaccattt ccgggtgcgg
     6841 gacggcgtta ccagaaactc agaaggttcg tccaaccaaa ccgactctga cggcagttta
     6901 cgagagagat gatagggtct gcttcagtaa gccagatgct acacaattag gcttgtacat
     6961 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     7021 taggtgacac tatagaatac acggaattaa
//
