LOCUS       sequence-244615-        6462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For AAV based expression of nuclear jRGECO1a in neurons in 
            Cre-negative neurons with FLEx switch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6462)
  AUTHORS   Farhi SL, Parot VJ, Grama A, Yamagata M, Abdelfattah AS, Adam Y, Lou
            S, Jun Kim J, Campbell RE, Cox DD, Cohen AE
  TITLE     Wide-area all-optical neurophysiology in acute brain slices.
  JOURNAL   J Neurosci. 2019 Apr 5. pii: JNEUROSCI.0168-19.2019. doi: 
            10.1523/JNEUROSCI.0168-19.2019.
  PUBMED    30952812
REFERENCE   2  (bases 1 to 6462)
  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: "J Neurosci.
            2019 Apr 5. pii: JNEUROSCI.0168-19.2019. doi: 
            10.1523/JNEUROSCI.0168-19.2019."
COMMENT     Sequence Label: SLF155: AAV-syn-DO-H2B-jRGECO1a
FEATURES             Location/Qualifiers
     source          1..6462
                     /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             791..1168
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1181..1190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1202..1258
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1954..1973
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    2447..2480
                     /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    2531..2564
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2589..3177
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2642..2662)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3060..3071)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3209..3685
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3408..3427)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3725..3865
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3725..3854
                     /label=AAV2 ITR
     rep_origin      3940..4395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4027..4046)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4237..4258
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4412..4431)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4531..4553
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4591..4609)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4677..4781
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4782..4850,4851..5642)
                     /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(5000..5019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5813..6401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6302..6321
                     /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 gcgccgcacc atgccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa
      841 ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag
      901 ctattccatc tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc
      961 caaggccatg ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggtga
     1021 ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac
     1081 ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa
     1141 ggccatcacc aagtacacca gcgctaagga tccaccggtc gccaccatgg tcgactcatc
     1201 acgtcgtaag tggaataagg caggtcacgc agtcagagct ataggtcggc tgagctcacc
     1261 cgtggtttcc gagcggatgt accccgagga cggcgccctg aagagcgaga tcaagaaggg
     1321 gctgaggctg aaggacggcg gccactacgc cgccgaggtc aagaccacct acaaggccaa
     1381 gaagcccgtg cagctgcccg gcgcctacat cgtggacatc aagttggaca tcgtgtccca
     1441 caacgaggac tacaccatcg tggaacagtg cgaacgcgcc gagggccgcc actccaccgg
     1501 cggcatggac gagctgtaca agggaggtac aggcgggagt ctggtgagca agggcgagga
     1561 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     1621 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg aggcctttca
     1681 gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     1741 ccctcagttc atgtacggct ccaaggccta cattaagcac ccagccgaca tccccgacta
     1801 cttcaagctg tccttccccg agggcttcag gtgggagcgc gtgatgaact tcgaggacgg
     1861 cggcattatt cacgtgaacc aggactcctc cctgcaggac ggcgtattca tctacaaggt
     1921 gaagctgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
     1981 ctgggaggct acgcgtgacg acctgactga agagcagatc gcagaattta aagaggcttt
     2041 ctccctattt gacaaggacg gggatgggac gataacaacc aaggagcttg ggacggtgtt
     2101 ccgatcgctg gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga
     2161 tgccgacggt gacggcacat tcgacttccc tgagttcctg acgatgatgg caagaaaaat
     2221 gaatgacaca gacagtgaag aggaaattcg cgaagcgttc cgcgtgtttg ataaggacgg
     2281 caatggctac atcggcgcag cagagcttcg ccacgtgatg acagaccttg gagagaagtt
     2341 aacagatgag gaggttgatg aaatgatcag ggtagcagac atcgatgggg atggtcaggt
     2401 aaactacgaa gagtttgtac aaatgatgac agcgaagtag gctagcataa cttcgtataa
     2461 agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat
     2521 ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc gatatcaagc
     2581 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2641 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2701 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2761 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2821 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2881 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2941 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3001 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3061 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3121 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3181 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3241 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3301 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3361 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     3421 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     3481 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     3541 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     3601 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     3661 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     3721 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3781 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3841 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3901 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3961 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4021 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4081 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4141 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4201 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4261 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4321 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4381 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4441 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4501 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4561 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4621 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4681 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4741 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4801 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4861 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4921 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4981 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5041 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5101 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5161 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5221 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5281 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5341 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5401 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5461 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5521 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5581 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5641 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5701 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5761 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5821 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5881 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5941 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6001 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6061 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6121 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6181 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6241 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6301 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6361 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6421 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
