LOCUS       sequence-432426-        6219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with hSynapsin promoter and DIO Lox sites for Cre-On 
            expression of red fluorescent calcium indicator jRGECO1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6219)
  AUTHORS   Meng C, Zhou J, Papaneri A, Peddada T, Xu K, Cui G
  TITLE     Spectrally Resolved Fiber Photometry for Multi-component Analysis of
            Brain Circuits.
  JOURNAL   Neuron. 2018 May 16;98(4):707-717.e4. doi: 
            10.1016/j.neuron.2018.04.012. Epub 2018 May 3.
  PUBMED    29731250
REFERENCE   2  (bases 1 to 6219)
  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: 
            "10.1016/j.neuron.2018.04.012"; journalName: "Neuron"; date: 
            "2018-05-16- 16"; volume: "98"; issue: "4"; pages: "707-717.e4"
COMMENT     Sequence Label: pAAV-hSyn-DIO-NES-jRGECO1a-WPRE
FEATURES             Location/Qualifiers
     source          1..6219
                     /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)."
     primer_bind     complement(1252..1271)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(1967..2023)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     CDS             complement(join(2051..2065,2066..2074))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(2078..2110)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(2114..2131)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2204..2237
                     /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    2288..2321
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2346..2934
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2399..2419)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2817..2828)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2966..3442
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3165..3184)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3482..3622
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3482..3611
                     /label=AAV2 ITR
     rep_origin      3697..4152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3784..3803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3994..4015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4169..4188)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4288..4310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4348..4366)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4434..4538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4539..4607,4608..5399)
                     /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(4757..4776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5570..6158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6059..6078
                     /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 gcgcctactt cgctgtcatc atttgtacaa actcttcgta gtttacctga ccatccccat
      841 cgatgtctgc taccctgatc atttcatcaa cctcctcatc tgttaacttc tctccaaggt
      901 ctgtcatcac gtggcgaagc tctgctgcgc cgatgtagcc attgccgtcc ttatcaaaca
      961 cgcggaacgc ttcgcgaatt tcctcttcac tgtctgtgtc attcattttt cttgccatca
     1021 tcgtcaggaa ctcagggaag tcgaatgtgc cgtcaccgtc ggcatctact tcattgatca
     1081 tgtcctgcag ctctgcttct gtggggttct gccccagcga tcggaacacc gtcccaagct
     1141 ccttggttgt tatcgtccca tccccgtcct tgtcaaatag ggagaaagcc tctttaaatt
     1201 ctgcgatctg ctcttcagtc aggtcgtcac gcgtagcctc ccagcccatg gtcttcttct
     1261 gcattacggg gccgtcgggg gggaagttgg tgccgcgcag cttcaccttg tagatgaata
     1321 cgccgtcctg cagggaggag tcctggttca cgtgaataat gccgccgtcc tcgaagttca
     1381 tcacgcgctc ccacctgaag ccctcgggga aggacagctt gaagtagtcg gggatgtcgg
     1441 ctgggtgctt aatgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg
     1501 cgaagggcag ggggccaccc ttggtcacct tcagcttagc ggtctgaaag gcctcgtagg
     1561 ggcggccctc gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgt
     1621 gcaccttgaa gcgcatgaac tccttgatga tggccatgtt atcctcctcg cccttgctca
     1681 ccagactccc gcctgtacct cccttgtaca gctcgtccat gccgccggtg gagtggcggc
     1741 cctcggcgcg ttcgcactgt tccacgatgg tgtagtcctc gttgtgggac acgatgtcca
     1801 acttgatgtc cacgatgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg
     1861 tcttgacctc ggcggcgtag tggccgccgt ccttcagcct cagccccttc ttgatctcgc
     1921 tcttcagggc gccgtcctcg gggtacatcc gctcggaaac cacgggtgag ctcagccgac
     1981 ctatagctct gactgcgtga cctgccttat tccacttacg acgtgatgag tcgaccattg
     2041 ttgcgagatc cttatcgtca tcgtcgtaca gatcccgacc catttgctgt ccaccagtca
     2101 tgctagccat accatgatga tgatgatgat gagaacctcc tccagtcttg ttaatatcaa
     2161 gtccagccaa cttaagagca agctcgttct gcagcatgct agcataactt cgtataaagt
     2221 atcctatacg aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt
     2281 gcaaagaata acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta
     2341 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2401 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2461 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2521 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2581 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2641 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2701 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     2761 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2821 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2881 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     2941 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     3001 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3061 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     3121 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     3181 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     3241 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     3301 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     3361 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3421 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     3481 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3541 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3601 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3661 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3721 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     3781 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3841 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3901 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3961 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4021 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4081 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4141 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4201 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4261 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4321 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4381 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4501 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4561 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4621 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4681 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4741 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4801 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4861 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4921 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4981 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5041 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5101 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5161 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5221 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5281 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5341 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5401 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5461 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5521 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5701 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5941 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6181 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
