LOCUS       sequence-479154-        6154 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Cre-dependent expression of axon targeted jGCaMP8s under hSyn 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6154)
  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 6154)
  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-Axon-jGCaMP8s
FEATURES             Location/Qualifiers
     source          1..6154
                     /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     1345..1364
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1604..1625
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1731..1752)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2034..2051)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2139..2172
                     /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    2223..2256
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2281..2869
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2334..2354)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2752..2763)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2901..3377
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3100..3119)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3417..3557
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3417..3546
                     /label=AAV2 ITR
     rep_origin      3632..4087
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3719..3738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3929..3950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4104..4123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4223..4245
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4283..4301)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4369..4473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4474..4542,4543..5334)
                     /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(4692..4711)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5505..6093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5994..6013
                     /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 gcgccttact tcgctgtcat catttgtaca aactcttcgt agtttacctg accatcccca
      841 tcgatgtctg cttccctgat catttcatca acctcttcat ctgttaactt ctctccaagg
      901 tttgtcatca cgtggcgaag ctctgctgca ctgatgtagc cattgccatc cttatcaaac
      961 acaccgaacg cttctctaat ttcttcttcc gtgtccctgt atttcatttt tcttgccatc
     1021 attgtcagga actcagggaa gtcgattgtg ccgtcaccgt cggcatctac ttcattgatc
     1081 atgtcctgca gctctgcttc tgtggggttg tgtccgagag accgcatcac cgtccccagc
     1141 tccttggttg ttattgtccc atccccgtcc ttgtcaaata gggagaaagc ctctttgtat
     1201 tctgcgatct gctcttcagt cagttggtcc ggcaggttgt actccagctt gtgccccagg
     1261 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
     1321 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
     1381 cgctcctgga tgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
     1441 tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag
     1501 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
     1561 tcgccctcac cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
     1621 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat gctccctccg
     1681 gtaccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     1741 agcaggacca tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga ctccacgctc
     1801 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     1861 tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc ggatgtggaa gttcgccttg
     1921 atgccgttct tctgcttgtc ggccttgata tagacgttga tcttcaggcc catcagcatg
     1981 gcgatgatct tcacggcggt ggccacctcc ttgaaggtct tcttgcgacg cgtgtgatga
     2041 tggtgatgat gcataatctt ttggtcctca tcattctttt caacctgttt ggttcttctc
     2101 atacagcaca gcatggtggc gggtggatcc ttgctagcat aacttcgtat aaagtatcct
     2161 atacgaagtt atttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa
     2221 gaataacttc gtataatgta tgctatacga agttatgaat tcgatatcaa gcttatcgat
     2281 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2341 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2401 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2461 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2521 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2581 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2641 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     2701 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     2761 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     2821 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     2881 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     2941 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     3001 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     3061 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     3121 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     3181 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     3241 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     3301 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     3361 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     3421 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3481 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3541 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3601 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3661 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3721 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3781 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3841 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3901 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3961 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     4021 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4081 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4141 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4201 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4261 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4321 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4381 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4441 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4501 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4681 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4741 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4801 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4861 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4921 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4981 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5041 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5101 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5161 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5221 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5281 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5341 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5401 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5461 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5521 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5581 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5641 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5701 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5761 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5821 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5881 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5941 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6001 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6061 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6121 ttcctggcct tttgctggcc ttttgctcac atgt
//
