LOCUS       sequence-379925-        6813 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV transfer plasmid for neuronal expression of Cre-dependent 
            (FLEX), soma-targeted (RiboL1) jGCaMP8s..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6813)
  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 6813)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-RiboL1-jGCaMP8s
FEATURES             Location/Qualifiers
     source          1..6813
                     /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     2074..2093
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2333..2354
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2460..2481)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2763..2780)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2798..2831
                     /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    2882..2915
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2940..3528
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2993..3013)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3411..3422)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3560..4036
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3759..3778)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4076..4216
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4076..4205
                     /label=AAV2 ITR
     rep_origin      4291..4746
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4378..4397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4588..4609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4763..4782)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4882..4904
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4942..4960)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5028..5132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5133..5201,5202..5993)
                     /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(5351..5370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6164..6752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6653..6672
                     /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 gcgccctaat acagacgctg gggcttgccc atggtgctct tgatgtacag agcccgcacg
      841 ttttgccagt ttttcttaag caaggacacc aagaaattga cagccagatg aatgttgtag
      901 actagctcat catcggtcat cttcacgtgg ccaacagcga cggccaaaca cagcaccttc
      961 ttcatctgga acttgattgt cgatttcacc tcatccactt tggccaccat gttttcattg
     1021 tgtgtcagca gggaggggaa cttgccagcc ttgtttaggc ctgggcccag gatacgtggg
     1081 atctgcttaa tcagagactc agaggccaaa aaggcatcgt acttcttagc cagcttcttg
     1141 accaacttct tgtttttgtt aagtttcttg agcgcctcga tgtccatgtg ggggatatcc
     1201 acggccttgg cttcatcaca gtgctgctgg tcccccagaa cgcacaccga gaacttgggg
     1261 cgtggggtgg acttgagcct gacggtgccc gagaaacgtt tgtccttctg agggtcgtag
     1321 ttcttcaggc tgatctgcag ctccaccgtc tccagaaact tgcggcgctt gcgctggttc
     1381 ccgtgcagga cttcccgcac cgcctcgtac agggtgtcgc gtgagacttt gctgcttgtt
     1441 ccaccactac cgccgcttcc ccctgtccca ccgctaccac cgctaccccc agtccctcca
     1501 gacccaccgc tgcccccctt cgctgtcatc atttgtacaa actcttcgta gtttacctga
     1561 ccatccccat cgatgtctgc ttccctgatc atttcatcaa cctcttcatc tgttaacttc
     1621 tctccaaggt ttgtcatcac gtggcgaagc tctgctgcac tgatgtagcc attgccatcc
     1681 ttatcaaaca caccgaacgc ttctctaatt tcttcttccg tgtccctgta tttcattttt
     1741 cttgccatca ttgtcaggaa ctcagggaag tcgattgtgc cgtcaccgtc ggcatctact
     1801 tcattgatca tgtcctgcag ctctgcttct gtggggttgt gtccgagaga ccgcatcacc
     1861 gtccccagct ccttggttgt tattgtccca tccccgtcct tgtcaaatag ggagaaagcc
     1921 tctttgtatt ctgcgatctg ctcttcagtc agttggtccg gcaggttgta ctccagcttg
     1981 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     2041 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     2101 aagatggtgc gctcctggat gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     2161 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     2221 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     2281 taggtggcat cgccctcacc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     2341 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     2401 ctccctccgg taccgccctt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     2461 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttcga aagtttggac
     2521 tccacgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     2581 tgctggtagt ggtaggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatgtggaag
     2641 ttcgccttga tgccgttctt ctgcttgtcg gccttgatat agacgttgat cttcaggccc
     2701 atcagcatgg cgatgatctt cacggcggtg gccacctcct tgaaggtctt cttgcgacgc
     2761 gtgtgatgat ggtgatgatg catggtggcg ggctagcata acttcgtata aagtatccta
     2821 tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag
     2881 aataacttcg tataatgtat gctatacgaa gttatgaatt cgatatcaag cttatcgata
     2941 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3001 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3061 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3121 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3181 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3241 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3301 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3361 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3421 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3481 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     3541 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     3601 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     3661 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     3721 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     3781 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     3841 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     3901 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     3961 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     4021 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     4081 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4141 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4201 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4261 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4321 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4381 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4441 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4501 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4561 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4621 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4681 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4741 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4801 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4861 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4921 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4981 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5041 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5101 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5161 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5221 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5281 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5341 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5401 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5461 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5521 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5581 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5641 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5701 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5761 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5821 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5881 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5941 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6001 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6061 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6121 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6181 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6241 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6301 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6361 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6421 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6481 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6541 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6601 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6661 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6721 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6781 tcctggcctt ttgctggcct tttgctcaca tgt
//
