LOCUS       sequence-361587-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre dependent co-expression of jGCaMP8s and soma-targeted ChrimsonR 
            under the control of an hSyn promoter. jGCaMP8s and ChrimsonR are 
            separated by cleavable peptide sequence P2A..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  AUTHORS   LaFosse PK, Zhou Z, Friedman NG, Deng Y, Li AJ, Akitake B, Histed MH
  TITLE     Bicistronic Expression of a High-Performance Calcium Indicator and 
            Opsin for All-Optical Stimulation and Imaging at Cellular 
            Resolution.
  JOURNAL   eNeuro. 2023 Mar 29;10(3):ENEURO.0378-22.2023. doi: 
            10.1523/ENEURO.0378-22.2023. Print 2023 Mar.
  PUBMED    36858826
REFERENCE   2  (bases 1 to 7420)
  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: 
            "10.1523/ENEURO.0378-22.2023"; journalName: "eNeuro"; date: 
            "2023-03-29- 29"; volume: "10"; issue: "3"; pages: 
            "ENEURO.0378-22.2023"
COMMENT     Sequence Label: pAAV-hSyn-DIO-jGCaMP8s-P2A-ChrimsonR-ST
FEATURES             Location/Qualifiers
     source          1..7420
                     /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(1005..2054)
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=ChrimsonR
                     /note="K176R mutant with accelerated closing kinetics"
                     /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
                     WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
                     AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
                     FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
                     SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
                     LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
                     FVEEEDEDTV"
     CDS             complement(2061..2117)
                     /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"
     primer_bind     2683..2702
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2942..2963
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3069..3090)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3372..3389)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    3405..3438
                     /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    3489..3522
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3547..4135
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3600..3620)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4018..4029)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4167..4643
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4366..4385)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4683..4823
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4683..4812
                     /label=AAV2 ITR
     rep_origin      4898..5353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4985..5004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5195..5216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5370..5389)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5489..5511
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5549..5567)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5635..5739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5740..5808,5809..6600)
                     /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(5958..5977)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6771..7359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7260..7279
                     /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 gcgccttaga atctggtggc ttcagggaag tccgtggcac agctgatgaa gctgtcaatg
      841 ctggacatgc tccgcatgtc gatgacgccc tccgtgcgtg cgggcagagg ggccacgggg
      901 ctgggcatgc tgctcatctc cagctcctct ggaggcttgc tctgcggggc catctccttg
      961 gtgttgagga tgggctggga ctgctctagg gctaccggtg ccgccactgt gtcctcgtcc
     1021 tcctcctcca cgaactcttc cacttccacc tcctcgcctc cgatctccat cttggtggtc
     1081 ttccggatgt cgccgtggat caggatgtgc tcgtggatct tgatcctcag gtggtgggcc
     1141 aggaaggtcc aaaactcctt ggcgatgatg tcgcagatgc tgtggccgat gctgttggcg
     1201 taagggctca gcttcagcag tccttctggt cccactgccc agaggattgg gtagctgccc
     1261 caagaggcga agtaagcgta ggccatcagc ttcacgacca tgcggcaatg gcctttaggc
     1321 acgctgtggt tggcttccac gtagcacttg gcggcctgga agtacatgta gccgccgtag
     1381 atgcaagaca cgatatacag cagccacttg agccaatcgg tagccagtcc tgcggccatg
     1441 ccgaacacga tcattcccac gcaagacacg atcaggccca tggtccgctt gctgtagtcg
     1501 ttcttcaggc cgctcaggtt gctcagtctg atcaggatca cggggcaaga cagcagccac
     1561 tcgaagtagc gcaggcaata ggcgtggttg ccggtagaca ggtacactgt ggcggggctg
     1621 ctgaactcct tgaagatctc cagggtcacg aacagcacct cgacgcaaca gacgtagacc
     1681 tcctcccaac cgcaagtggc cttccaggcg ctgaagccgt agaatgtcag cagggcgatg
     1741 gcgatgctga aagcaatcca ctggcagacc tgggcgccga tcttttctcc tggtgtgcct
     1801 ctgctgtgca gccaagcctg cttcttctcc actcccacga ccaggtagcc gcctgtggcc
     1861 tccacgaagc aatatccgta tccggcgtcg ctcagtccgt aagaagggtc acaccaccac
     1921 tcggtgctga agcactcgcc tgtaggtgtc attcctccac agcccatgtc ctcgtggtgg
     1981 taggggttag gccaagggtt gatgcctccg gcggcaaaca gagatctggt ggcgctgctg
     2041 atcagctcag ccatactagt aggtccaggg ttctcctcca cgtctccagc ctgcttcagc
     2101 aggctgaagt tagtagctcc gcttcccttc gctgtcatca tttgtacaaa ctcttcgtag
     2161 tttacctgac catccccatc gatgtctgct tccctgatca tttcatcaac ctcttcatct
     2221 gttaacttct ctccaaggtt tgtcatcacg tggcgaagct ctgctgcact gatgtagcca
     2281 ttgccatcct tatcaaacac accgaacgct tctctaattt cttcttccgt gtccctgtat
     2341 ttcatttttc ttgccatcat tgtcaggaac tcagggaagt cgattgtgcc gtcaccgtcg
     2401 gcatctactt cattgatcat gtcctgcagc tctgcttctg tggggttgtg tccgagagac
     2461 cgcatcaccg tccccagctc cttggttgtt attgtcccat ccccgtcctt gtcaaatagg
     2521 gagaaagcct ctttgtattc tgcgatctgc tcttcagtca gttggtccgg caggttgtac
     2581 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
     2641 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     2701 tccttgaaga agatggtgcg ctcctggatg tagccttcgg gcatggcgga cttgaagaag
     2761 tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg
     2821 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc
     2881 agcttgccgt aggtggcatc gccctcaccc tcgccggaca cgctgaactt gtggccgttt
     2941 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     3001 ctcaccatgc tccctccggt accgcccttg tacagctcgt ccatgccgag agtgatcccg
     3061 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttcgaa
     3121 agtttggact ccacgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     3181 ggggtgttct gctggtagtg gtaggcgagc tgcacgccgc cgtcctcgat gttgtggcgg
     3241 atgtggaagt tcgccttgat gccgttcttc tgcttgtcgg ccttgatata gacgttgatc
     3301 ttcaggccca tcagcatggc gatgatcttc acggcggtgg ccacctcctt gaaggtcttc
     3361 ttgcgacgcg tgtgatgatg gtgatgatgc atggtggcgc tagcataact tcgtataaag
     3421 tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg
     3481 tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgaattcga tatcaagctt
     3541 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3601 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3661 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3721 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3781 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3841 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3901 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     3961 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4021 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4081 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     4141 taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc
     4201 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     4261 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     4321 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     4381 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     4441 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     4501 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     4561 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     4621 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc
     4681 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4741 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     4801 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     4861 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4921 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     4981 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5041 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5101 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5161 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5221 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     5281 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5341 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5401 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5461 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5521 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5581 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5701 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5761 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5821 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5881 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5941 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6001 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6061 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6121 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6181 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6241 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6301 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6361 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6421 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6481 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6541 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6601 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6661 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6721 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6781 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6841 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6901 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6961 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7021 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7081 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7141 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7201 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7261 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7321 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7381 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
