LOCUS       sequence-478701-        6434 bp    DNA     circular SYN 15-JUL-2025
DEFINITION  AAV-mediated expression of positive-going voltage sensor under the 
            Syn promoter, Cre-dependent expression; soma localization targeting 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6434)
  AUTHORS   Eric R. Schreiter, Ahmed Abdelfattah, Jeremy Hasseman, Daniel Reep, 
            Getahun Tsegaye, Arthur Tsang, Ilya Kolb, Allan Wong, Amy Chuong, 
            Jihong Zheng, Ben Arthur, Wyatt Korff, GENIE Project Team
  TITLE     Optimization of Ace2N based voltage indicators
  JOURNAL   Janelia Figshare 2023
REFERENCE   2  (bases 1 to 6434)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Janelia 
            Figshare 2023"
COMMENT     Sequence Label: pGP-AAV-syn-FLEX-Positron2-ST-WPRE
FEATURES             Location/Qualifiers
     source          1..6434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     promoter        256..703
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    733..766
                     /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    829..862
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2723..2732
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2743..2776)
                     /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(2847..2880)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2922..3510
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2975..2995)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3393..3404)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3553..3674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3564..3583)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3618..3637
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3836..3965
                     /label=AAV2 ITR
     rep_origin      complement(4202..4790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4282..4301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4961..5752,5753..5821))
                     /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     5584..5603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5822..5926)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5952..6407)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6089..6110)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6301..6320
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc
       61 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
      181 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag atctctgcag
      241 agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc
      301 ctacctgacg accgaccccg acccactgga caagcaccca acccccattc cccaaattgc
      361 gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc
      421 agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct
      481 cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca
      541 ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat
      601 aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc
      661 tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt caagctgcta
      721 gcaaggatcc ccataacttc gtataaagta tcctatacga agttatatca aaataggaag
      781 accaatgctt caccatcgac ccgaattgcc aagcatcacc atcgacccat aacttcgtat
      841 aatgtatgct atacgaagtt atactagagc tttcagaatc gagtagcctc ggggaaatct
      901 gttgcgcaag agatgaagct atcaatagat gacatactcc tcatgtcaat cactccttca
      961 gtcctggcag ggaggggagc cacgggactt ggcatactgg acatctcaag ctcttcggga
     1021 ggcttagact gtggagccat ttcctttgtg ttcaggatag gctgactttg cacctcgttc
     1081 tcgtagcaga acacgttgat gtcgatctgg tccaggggga tgtactcgcc ctcgctggtg
     1141 atcctgctct tagtggttgg ctcgccggaa atctcgagcg tcgacagcca gcgcgcgatc
     1201 tcgctgccga tcaggtccgg gttgtcttct tgcagcagat tcagacccgg gccgatgtcc
     1261 acagccttgc agttaggcag gcttttggcc aggcgagcgg cttcggccgg tgggatcaga
     1321 acgcctgggg tgccccagaa cagcagcttc gggacagggg actggtgcag ccagtccatg
     1381 tattcttcga ccagcgcgac gatgttcgct ggctcaccgg cgattggcag ctcgtttggg
     1441 aagcgccaca gtggctcgcg gtcaacagga ttcaggaacg gctcgcggta atggtccatc
     1501 tcgacttcag tcagcgggcg gacgacaccc atcggcagcg taccctcgat aaaaacgttc
     1561 tgatcgatga tcagcttgcg gccgacgtcg gtggtgcgga aggcctggaa ggtctcgcgg
     1621 gcaaattctg gccattcgtc ccaggtcggg atagggcgga tgaactccat aaatgcaata
     1681 cctttgacgc gctctggatt gcgcttggcc cagtggaaac ccagagcgga gccccagtcg
     1741 tgaatgacca ggacgacctc ttccagaccc agggcttcga tgaaggcatc catgaagcgg
     1801 acgtggtcgt cgaagaaata acccaggtct ggtttgtcgg atttgcccat accgatcagg
     1861 tctggagcaa tgcagcgatg ggtcggtgca acatgcggga tgatgttgcg ccacacgtag
     1921 gaggaggtcg ggttaccgtg caggaacagc acaggggtgc catcgcgcgg accaacatcg
     1981 acgtagtgca tgcgctcgcc caggacttcc acataatggg ggtcgaatgg aaagccggta
     2041 ccgatcttga agatagtctc atgggcaatg aggcacccca ttccaaacag tggcttagcg
     2101 agcacatcca ggattcccat gagaatggcc tcattgtcga cggacagcac gccactacca
     2161 gacccgaaag cccacacgac aggataaaac acgaagagga ttgacaggaa gacgagcaga
     2221 ctgacatagg agctatggta ctcggggctt ttcttcagat cgtctttcca agtggcgtta
     2281 tacatcccgt atgtcaggac tgcctgcatc acacatccag cgataaagta cacccatttg
     2341 aacttgtgtt catcctcgaa ggcgcccaga ataccaaaca cgatcatgaa aatgtctgcg
     2401 ccgatgaccc aagaaatcat cactccgccc atcttggtca tgacgatgag gttgagcagg
     2461 agcagtggtg tggtcagcac ccagtcaata tattttgcgt agaccacctg cctttcacca
     2521 ttcattgtca ggtcctggta gttcacggcc attgcatagt aagcggtagc ggcgatagtg
     2581 cagatggcaa tgttgatata gtatgcgctc ttcagctctg aaggccgaaa gtgcacagcc
     2641 actccaaaag caatagcagc agcagccata atagcaaaga caatccactg tgcaatcata
     2701 ccggtctcgg tttccacgtc agccatggtg gcggatccta gtataacttc gtataggata
     2761 ctttatacga agttatcatt gggattcttc ctattttgat ccaagcatca ccatcgaccc
     2821 tctagtccag atctcaccat cgacccataa cttcgtatag catacattat acgaagttat
     2881 gtccctcgaa gaggttcgaa ttcgatatca agcttatcga taatcaacct ctggattaca
     2941 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3001 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3061 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3121 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3181 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3241 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3301 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     3361 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3421 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3481 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacccggg cggccgcttc
     3541 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     3601 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     3661 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     3721 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     3781 ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa
     3841 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3901 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3961 cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4021 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4081 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4141 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4621 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5941 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     6001 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     6061 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     6121 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     6181 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     6241 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     6301 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     6361 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     6421 caggctgcgc aact
//
