LOCUS       sequence-441133-        6204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuron-specific, Cre-dependent co-expression of FLAG-tagged 
            inhibitory KORD DREADD receptor, and a single-cell discriminating 
            version of AausFP1 as a reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6204)
  AUTHORS   Szelenyi ER, Navarrete JS, Murry AD, Zhang Y, Girven KS, Kuo L, 
            Cline MM, Bernstein MX, Burdyniuk M, Bowler B, Goodwin NL, Juarez B,
            Zweifel LS, Golden SA
  TITLE     An arginine-rich nuclear localization signal (ArgiNLS) strategy for 
            streamlined image segmentation of single cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Aug 6;121(32):e2320250121. doi: 
            10.1073/pnas.2320250121. Epub 2024 Jul 29.
  PUBMED    39074275
REFERENCE   2  (bases 1 to 6204)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-08-6"; volume: "121(32):e2320250121. 
            doi"; pages: " 10.1073/pnas.2320250121. Epub 2024 Jul 29"
COMMENT     Sequence Label: pAAV-hSyn-DIO-FLAG-KORD-P2A-ArgiNLS-AausFP1
FEATURES             Location/Qualifiers
     source          1..6204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..592
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(611..644)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(695..728)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1532..1588)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2093..2104
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(2813..2836)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2836..2845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2852..2885
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2936..2969
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3066..3086)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3468..3608
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3468..3597
                     /label=AAV2 ITR
     rep_origin      3683..4138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3770..3789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3980..4001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4155..4174)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4274..4296
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4334..4352)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4420..4524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4525..4593,4594..5385)
                     /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(4743..4762)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5556..6143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6044..6063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagtgca agtgggtttt aggaccagga tgaggcgggg
      181 tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc
      241 ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg
      301 cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca
      361 ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct
      421 tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag
      481 gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct
      541 gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agtctagagt
      601 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
      661 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
      721 gaagttatgg cgcgccttag gcgtaggcgg tcttgcagtc gatggctctg aacacttcca
      781 ccacgttcat gtggtcccgg gtttctttag ggtccttgct caggatggtg tgggcctgaa
      841 tgtggtgata gtgagggatg tccacggggc cacctccgag aggggtattc agctgctggt
      901 gagcagcaat gatctggcct ccgccgatca cgtcgtgcac aatgttgatg tagcacttga
      961 tgccgttgtt ctgcacgtcg ggcagcacgt agatgcagtg cgggttgtag ttgaactcaa
     1021 gtttcttgcc caggatgttg ccgcctttct tgaagccgct gccgttcagg gtcactctgt
     1081 tgtacacgga gcccttctcg taggtcactt cggctctggt cttgtacacg ccgtcgttct
     1141 cgaaggtgat ggttctctct tgcacgtagc cctcaggcat ggcgctcttg gggtagtcct
     1201 tgatgtcgtt gggatacttg gcgaagcact gggcgccgta tgtcactgtg gtcaggatgc
     1261 tgctccaagg cacaggcagt tcgccggttg tgcagatgaa gctggcctcg atctttccgg
     1321 tattggcgtc gccgtgtcct ttgcctctga cgctgaactt catgccttcc acgtcgcctt
     1381 ccatttccac cacatagggg atcttctcgc ggaacagcag ggcgccataa gacatggtgg
     1441 ccacagggga tcccgttcga cggcgccgcc gccttctcac cttctttttc tttgctatgc
     1501 tcatctgctt ctggggagcc acccttttca taggaccggg gttttcttcc acgtctcctg
     1561 cttgctttaa cagagagaag ttcgtggctc cggatcctac tggtttattc atcccatcga
     1621 tgtccctcag gtaagcagga tcctgaactg tatttcggac tctgctagtg ctctgccgct
     1681 ccatcctcat cttcagtgga aagcagaagt cccggaaaca ccgcttgaag ttttcatcaa
     1741 gaaaggcgta gagaatggga ttcaggctac tgttggtata gcctaaggcg atgcagaagt
     1801 aatagctgga gagagcagct gtgctgtggg aggtgctccc cagagcctcc accaggatga
     1861 atatgtgaat gggagtccag cagacgacga agactgccac caccaccagg accagtctgg
     1921 tgatcctacg caggttgcga tctttctctc gggagccaga aaggagccgg acgctcttga
     1981 gacgcaggat catcagggtg tagcagacga tgatgatgag gacagggatc acgaaggcaa
     2041 agatgaagac gcagatcttc atgaagaggt cccaccagga gtagtcatca tctgggaact
     2101 gcaaggagca ctcaatgaca tcgacgtctt ccctgacttt ggtgcctcca aggactattg
     2161 cagagatgcc aacagatgac gacagcagcc agatgcagat attgatgatc tttgccttca
     2221 agggtgtgcg gaagtccaaa gccttcacgg ggtggcacac ggcaatgtag cggtccacgc
     2281 tcatcatggt caaggtgaag atgctggtga acatgttgta gtaattaatg gaaattacta
     2341 tcttgcacag cacatcccca aaaggccagg aattcatcaa gtagaccgta ctctgaaagg
     2401 gcatggttgt agtaactaaa gcatctgcca aagccaggtt aaatatgtaa atgttggttg
     2461 ctgtcttcat ctttgtgtat cggatgatca cgaacatgac cagcgagttg cccaccaagc
     2521 ccacgacgaa cactacggag tagaccgccg tgatgatgac cgggatggcc ggggagatgt
     2581 gcgcgggctc cagctgcgcg tcctccgagc cggcgctgcc gttgctgtcg ggctcggccc
     2641 agccgggaaa ccaggcgctg ctgttggggg gcaggcaggc gctcggggcg caggtagggc
     2701 ccggctcccc gcggaagatc tgaatcgggg attcatttaa atcttcttct gataataatt
     2761 tttgttctaa tgcatctcga gtggatccga gctctgaatt ctcaccgata tctttatcat
     2821 catcatcttt ataatccatg gtggcgctag cataacttcg tataaagtat cctatacgaa
     2881 gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc aaagaataac
     2941 ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc gatatcgaat
     3001 tccgctcgag ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3061 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3121 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     3181 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3241 actgacaatt ccgtggtgtt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3301 atctagcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3361 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     3421 gtgggaggtt ttttaaagcg gccgccacgt gcggaccgag cggccgcagg aacccctagt
     3481 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3541 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3601 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3661 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3721 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     3781 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3841 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     3901 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3961 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4021 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4081 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4141 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4201 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4261 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4321 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4381 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4441 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4501 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4561 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4621 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4681 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4741 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4801 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4861 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4921 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4981 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5041 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5101 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5161 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5221 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5281 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5341 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5401 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5461 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5521 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5581 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5641 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5701 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5761 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5821 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5881 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5941 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6001 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6061 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6121 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6181 tttgctggcc ttttgctcac atgt
//
