LOCUS       sequence-432512-        6153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An axon-targeted jGCaMP8m controlled by hSynapsin1 promoter and flex
            switch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6153)
  AUTHORS   Abe K, Kambe Y, Majima K, Hu Z, Ohtake M, Momennezhad A, Izumi H, 
            Tanaka T, Matunis A, Stacy E, Itokazu T, Sato TR, Sato TK
  TITLE     Enhanced Aversive Signals During Classical Conditioning in Dopamine 
            Axons in Medial Prefrontal Cortex.
  JOURNAL   bioRxiv. 2023 Aug 24:2023.08.23.554475. doi: 
            10.1101/2023.08.23.554475. Preprint.
  PUBMED    37662305
REFERENCE   2  (bases 1 to 6153)
  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: "bioRxiv. 
            2023 Aug 24:2023.08.23.554475. doi: 10.1101/2023.08.23.554475. 
            Preprint."
COMMENT     Sequence Label: pAAV-hSynapsin1-FLEx-axon-jGCaMP8m-WPRE-SV40
FEATURES             Location/Qualifiers
     source          1..6153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   61..190
                     /label=AAV2 ITR
     rep_origin      262..717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(349..368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     559..580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        743..847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(848..916,917..1708)
                     /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(1066..1085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1879..2467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2368..2387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2704..2833
                     /label=AAV2 ITR
     polyA_signal    2995..3116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3032..3051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3086..3105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3194..3782
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3300..3311
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3709..3729
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3824..3857
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3928..3961
                     /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)."
     CDS             4062..4079
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     4361..4382
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4488..4509)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4749..4768)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(5347..5380)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(5443..5476)
                     /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)."
     primer_bind     complement(5487..5506)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5489..5621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
ORIGIN
        1 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
       61 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      121 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      181 gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      241 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      301 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      361 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      421 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      481 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      541 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      601 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      661 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
      721 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      781 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      841 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      901 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      961 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1021 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1081 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1141 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1201 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1261 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1321 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1381 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1441 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1501 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1561 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1621 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1681 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1741 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1801 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1861 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1921 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1981 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2041 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2101 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2161 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2221 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2281 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2341 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2401 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2461 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2521 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2581 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2641 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2701 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     2761 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2821 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
     2881 tctaggaaga tccttatcga ttttaccaca tttgtagagg ttttacttgc tttaaaaaac
     2941 ctcccacatc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3001 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3061 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3121 gtctgctcga agcggccgcc cgggtcgact ctagatgcat gctcgagcgg ccgcccgggt
     3181 cgacggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg
     3241 cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga
     3301 ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag
     3361 gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc
     3421 aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca
     3481 atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa
     3541 ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc
     3601 cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc
     3661 atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa
     3721 ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga
     3781 ttatcgataa gcttgatatc gaattcgaac ctcttcgagg gacataactt cgtataatgt
     3841 atgctatacg aagttatggg tcgatggtga gatctggact agagggtcga tggtgatgct
     3901 tggatcaaaa taggaagaat cccaatgata acttcgtata aagtatccta tacgaagtta
     3961 tactagaatt cggtaccgct agcactagtg ccaccatgct gtgctgtatg agaagaacca
     4021 aacaggttga aaagaatgat gaggaccaaa agattggttc tcatcatcac catcatcaca
     4081 cgcgtcgcaa gaagaccttc aaggaggtgg ccaccgccgt gaagatcatc ggccgcctga
     4141 tgggcctgaa gatcaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga
     4201 acttccacat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc
     4261 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgg
     4321 agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     4381 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga
     4441 gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     4501 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct
     4561 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     4621 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     4681 agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct
     4741 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4801 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4861 acaagctgga gtacaacctg ccggaccaac tgactgaaga gcagatcgca gaatacaaag
     4921 aggctttctc cctatttgac aaggacgggg atgggacaat aacaaccaag gagctgggga
     4981 cggtgatgcg gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg
     5041 aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcaa
     5101 gaaaaatgaa atacagggac acggaagaag aaattagaga agcgttcggt gtgtttgata
     5161 aggatggcaa tggctacatc agtgcagcag agcttcgcca cgtgatgaca aaccttggag
     5221 agaagttaac agatgaagag gttgatgaaa tgatcaggga agcagacatc gatggggatg
     5281 gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaagtgaggc gcgccgagct
     5341 cctagtataa cttcgtatag catacattat acgaagttat gtgtcgatgg tgatgcttgg
     5401 caattcgggt cgatggtgaa gcattggtct tcctattttg atataacttc gtataggata
     5461 ctttatacga agttatgggg atcatcacct gtggagagaa aggcaaagtg gatgtcagta
     5521 agaccaatag gtgcctatca gaaacgcaag agtcttctct gtctcgacaa gcccagtttc
     5581 tattggtctc cttaaacctg tcttgtaacc ttgatactta cctgatccaa cggctgcgac
     5641 ttggggcagg gggtcctagg ggtggtctgg ccaggagccg cgggggcgga ctgcgcggtg
     5701 cccaggagca cagctgcgct ctcaggcacg acacgactcc tccgctgccc accgcagact
     5761 gaggcagcgc tgagtcgccg gcgccgcagc gcagatggtc gcgcccgtgc ccccctatct
     5821 cgcgcctcgc gtggtgcggt ccggctgggc cggcggcggc gcggacgcga ccaaggtggc
     5881 cgggaagggg agtttgcggg ggaccggcga gtgacgtcag cgcgccttca gtgctgaggc
     5941 ggcggtggcg cgcgccgcca ggcgggggcg aaggcactgt ccgcggtgct gaagctggca
     6001 gtgcgcacgc gcctcgccgc atcctgtttc ccctccccct ctctgatagg ggatgcgcaa
     6061 tttggggaat gggggttggg tgcttgtcca gtgggtcggg gtcggtcgtc aatttaaata
     6121 ctgaaaaggg cgaattccag cacactggag ctc
//
