LOCUS       sequence-479152-        6435 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Cre-dependent expression of Soma-jGCaMP8f under hSyn promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6435)
  AUTHORS   Grodem S, Nymoen I, Vatne GH, Rogge FS, Bjornsdottir V, Lensjo KK, 
            Fyhn M
  TITLE     An updated suite of viral vectors for in vivo calcium imaging using 
            intracerebral and retro-orbital injections in male mice.
  JOURNAL   Nat Commun. 2023 Feb 4;14(1):608. doi: 10.1038/s41467-023-36324-3.
  PUBMED    36739289
REFERENCE   2  (bases 1 to 6435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-02-4"; volume: "14(1):608. doi"; pages: " 
            10.1038/s41467-023-36324-3"
COMMENT     Sequence Label: pAAV hSyn:FLEX-Soma-jGCaMP8f
FEATURES             Location/Qualifiers
     source          1..6435
                     /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     1696..1715
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1955..1976
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2082..2103)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2385..2402)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2420..2453
                     /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    2504..2537
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2562..3150
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2615..2635)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3033..3044)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3182..3658
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3381..3400)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3698..3838
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3698..3827
                     /label=AAV2 ITR
     rep_origin      3913..4368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4000..4019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4210..4231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4385..4404)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4504..4526
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4564..4582)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4650..4754
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4755..4823,4824..5615)
                     /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(4973..4992)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5786..6374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6275..6294
                     /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 gaggtccgta gcgtgtctcg tactggctca cgatgttgcg cagccgctgc
      841 actctctgcc tcagctcagc cacccttgtc ctcagggcgg tgttccgtct cctcagaaaa
      901 gcggcccgga tctccaggga tcccagtggt ccgtatctgg tctcgtactg agacacgatg
      961 ttctccagcc gctgcacctc ctgctccagc tcagccacct ctgtctccag ggcggtgttc
     1021 tcctgctcca ggaaagcggc ctcgatttcc agtgtcccac cgctaccgcc gcttccccct
     1081 gtcccaccgc taccaccgct acccccagtc cctccagacc caccgctgcc cccgagctcc
     1141 ttcgctgtca tcatttgtac aaactcttcg tagtttacct gaccatcccc atcgatgtct
     1201 gcttccctga tcatttcatc aacctcttca tctgttaact tctctccaag gtttgtcatc
     1261 acgtggcgaa gctctgctgc actgatgtag ccattgccat ccttatcaaa cacaccgaac
     1321 gcttctctaa tttcttcttc cgtgtccctg tatttcattt ttcttgccat cattgtcagg
     1381 aactcaggga agtcgattgt gccgtcaccg tcggcatcta cttcattgat catgtcctgc
     1441 agctctgctt ctgtggggtt cagtccgaga gaccgcatca ccgtccccag ctccttggtt
     1501 gttattgtcc catccccgtc cttgtcaaat agggagaaag cctctttaaa ttctgcgatc
     1561 tgctcttcag tcagttggtc cggcaggttg tactccagct tgtgccccag gatgttgccg
     1621 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     1681 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     1741 atgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     1801 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
     1861 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctca
     1921 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     1981 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tgctccctcc ggtaccgccc
     2041 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     2101 atgtgatcgc gcttctcgtt ggggtctttc gaaagtttgg actccacgct caggtagtgg
     2161 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtaggcg
     2221 agctgcacgc cgccgtcctc gatgttgtgg cggatgtgga agttcgcctt gatgccgttc
     2281 ttctgcttgt cggccttgat atagacgttg atcttcaggc ccatcaggcg ggcgatgatc
     2341 ttcacggcgg tggccacctc cttgaaggtc ttcttgcgac gcgtgtgatg atggtgatga
     2401 tgcatggtgg cgggctagca taacttcgta taaagtatcc tatacgaagt tatttgcctt
     2461 aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt
     2521 atgctatacg aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca
     2581 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2641 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2701 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2761 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2821 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2881 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2941 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     3001 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3061 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3121 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     3181 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     3241 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     3301 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     3361 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     3421 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     3481 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     3541 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     3601 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     3661 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3721 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3781 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3841 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3901 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3961 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     4021 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4081 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4141 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4201 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     4261 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     4321 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4381 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4441 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4501 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4561 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4621 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4681 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4741 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4801 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4861 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4921 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4981 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5041 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5101 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5161 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5221 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5281 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5341 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5401 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5461 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5521 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5581 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5641 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5701 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5761 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5821 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5881 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5941 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6001 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6061 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6121 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6181 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6241 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6301 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6361 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6421 cttttgctca catgt
//
