LOCUS       sequence-377587-        6279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-on expression of soma-targeted QuasAr6a under an neuronal 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6279)
  AUTHORS   Tian H, Davis HC, Wong-Campos JD, Park P, Fan LZ, Gmeiner B, Begum 
            S, Werley CA, Borja GB, Upadhyay H, Shah H, Jacques J, Qi Y, Parot 
            V, Deisseroth K, Cohen AE
  TITLE     Video-based pooled screening yields improved far-red genetically 
            encoded voltage indicators.
  JOURNAL   Nat Methods. 2023 Jul;20(7):1082-1094. doi: 
            10.1038/s41592-022-01743-5. Epub 2023 Jan 9.
  PUBMED    36624211
REFERENCE   2  (bases 1 to 6279)
  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"; doi: 
            "10.1038/s41592-022-01743-5"; journalName: "Nat Methods"; date: 
            "2023-07"; volume: "20"; issue: "7"; pages: "1082-1094"
COMMENT     Sequence Label: HT109_pAAV_hSyn-DiO-SomQuasAr6a_EGFP
FEATURES             Location/Qualifiers
     source          1..6279
                     /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        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /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(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1029..1739,1740..1742))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(1029..1739,1740..1742))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1071..1092)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1420..1439
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1679..1700
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2606..2615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2626..2659
                     /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    2710..2743
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2768..3356
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2821..2841)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3239..3250)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3410..3429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3464..3483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3542..3682
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3542..3671
                     /label=AAV2 ITR
     rep_origin      3757..4212
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3844..3863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4054..4075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4229..4248)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4348..4370
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4408..4426)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4494..4598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4599..4667,4668..5459)
                     /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(4817..4836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5630..6218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6119..6138
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccttaca cctcgttctc gtagcagaac ttgtagaacc tggtggcctc agggaagtcg
      841 gtggcgcagc tgatgaagct gtcgatgctg ctcatgctcc tcatgtcgat cacgccctcg
      901 gtcctggcag gcagaggggc cacagggcta ggcatgctgc tcatctccag ctcctcagga
      961 ggcttgctct gaggggccat ctccttggtg ttcaggatag gctggctctg ggtacctccg
     1021 ctcccaccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     1081 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc
     1141 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     1201 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     1261 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     1321 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     1381 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     1441 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     1501 tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc
     1561 acgagggtgg gccagggcac gggcagcttg ccggtggtgc aaatgaactt cagggtcagc
     1621 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     1681 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     1741 accacgttga tgtcgatctg gtccaggggg atgtactcgc cctcgctggt gatcctgctc
     1801 ttggcggccg ccttgctcac tgctactacc gggcggtcgg cggcaccggt atcagctcca
     1861 gctgaaggtt caggtgcttc tgtatcttgc agaatgcttc ttgacctgag caggataaaa
     1921 ccgaagccca ccttggcggt gacgtccagc accatgaaca gcaaagtttc gattcccagg
     1981 cccacaactc cggcaccctc ggtgccgata atccagatga tagggtaagc agtccacagg
     2041 accagaacca gtgcggtgag gatgttgaag gttgaggcaa cctctgggcc tctctcctta
     2101 gcggctgacc gcagagaagt ggccaggaca tacagaacga cgatcataca gatagttgag
     2161 aacaggcacc aagagtatct ggcaatggca gtatgtgaca gtgtccctat caggcctgtg
     2221 acaatcatca gagcatcgcc gccaacgaga gtgccgatgg tcactctatc aactttggcc
     2281 aggagtgcca ggtgcagcag caacagtggt gtagtcagca gccactgtgc gtatcttgcg
     2341 tagtagatgt tcaggatctc tccacctact ggcacctctg tgaggccaat accaaagaac
     2401 attgacagat aagctgcact ggcaataccg ctgaccagga ttgtcactgc gtagtactcc
     2461 cgtgcatcct tgtcagtctc tccgcccgct ctaaccagga agtagaaggt tccgatcagc
     2521 atcagcagtg taccaattcc gagccacaga ctttcaggtc taccatctcc gaggaggtca
     2581 taaccagcct gcagtgcgat agaaaccatg gtggcggagg ctagcataac ttcgtataaa
     2641 gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg
     2701 gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg atatcaagct
     2761 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2821 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2881 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2941 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3001 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3061 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3121 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3181 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3241 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3301 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3361 ataccgtcga cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3421 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3481 tgctttattt gtaaccatta taagctgcaa taaactaacc acgtgcggac cgagcggccg
     3541 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3601 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3661 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3721 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3781 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     3841 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3901 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3961 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4021 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4081 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4141 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4201 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4261 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4321 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4381 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4441 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4501 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4561 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4621 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4681 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4741 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4801 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4861 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4921 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4981 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5041 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5101 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5161 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5221 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5281 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5341 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5401 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5461 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5521 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5581 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5641 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5701 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5761 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5821 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5881 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5941 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6001 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6061 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6121 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6181 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6241 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
