LOCUS       sequence-399228-        5532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For enriched expression of GCaMP6s in axons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5532)
  AUTHORS   Broussard GJ, Liang Y, Fridman M, Unger EK, Meng G, Xiao X, Ji N, 
            Petreanu L, Tian L
  TITLE     In vivo measurement of afferent activity with axon-specific calcium 
            imaging.
  JOURNAL   Nat Neurosci. 2018 Sep;21(9):1272-1280. doi: 
            10.1038/s41593-018-0211-4. Epub 2018 Aug 20.
  PUBMED    30127424
REFERENCE   2  (bases 1 to 5532)
  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: 
            "10.1038/s41593-018-0211-4"; journalName: "Nat Neurosci"; date: 
            "2018-09"; volume: "21"; issue: "9"; pages: "1272-1280"
COMMENT     Sequence Label: pAAV-hSynapsin1-axon-GCaMP6s
FEATURES             Location/Qualifiers
     source          1..5532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      200..655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(287..306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     497..518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        681..785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(786..854,855..1646)
                     /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(1004..1023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1817..2405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2306..2325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2784..2905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2821..2840)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2875..2894
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2934..2950
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2935..2951
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2954..3542
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3060..3071
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3469..3489
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3555..3998,3999..4004,4005..4439,4440..4457,
                     4458..4727,4728..4733,4734..4790,4791..4817,4818..4841,
                     4842..4844,4845..4877,4878..4880,4881..4898,4899..4904))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="GSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNKT
                     GHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSL
                     GQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGNG
                     YISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     4114..4133
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4373..4394
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4500..4521)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      4802..4811
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5002..5449)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   join(5531..5532,1..128)
                     /label=AAV2 ITR
ORIGIN
        1 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
       61 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
      121 gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
      181 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      241 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      301 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      361 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      421 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      481 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      541 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      601 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
      661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      781 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      841 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      901 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      961 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1021 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1081 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1141 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1201 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1261 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1321 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1381 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1441 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1501 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1561 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1621 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1681 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2461 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2521 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgagcgag
     2581 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc
     2641 cgccatgcta cttatctacg tagccatgct ctaggaagat ccttatcgat tttaccacat
     2701 ttgtagaggt tttacttgct ttaaaaaacc tcccacatct ccccctgaac ctgaaacata
     2761 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2821 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2881 tgtccaaact catcaatgta tcttatcatg tctgctcgaa gcggccgccc gggtcgaggt
     2941 cgacggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg
     3001 cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga
     3061 ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag
     3121 gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc
     3181 aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca
     3241 atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa
     3301 ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc
     3361 cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc
     3421 atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa
     3481 ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga
     3541 ttatcgataa gcttttactt cgctgtcatc atttgtacaa actcttcgta gtttacctga
     3601 ccatccccat cgatgtctgc ttccctgatc atttcatcaa cctcttcatc tgttaacttc
     3661 tctccaaggt ttgtcatcac gtggcgaagc tctgctgcac tgatgtagcc attgccatcc
     3721 ttatcaaaca caccgaacgc ttctctaatt tcttcttccg tgtccctgta tttcattttt
     3781 cttgccatca ttgtcaggaa ctcagggaag tcgattgtgc cgtcaccgtc ggcatctact
     3841 tcattgatca tgtcctgcag ctctgcttct gtggggttct gccccagaga ccgcatcacc
     3901 gtccccagct ccttggttgt tattgtccca tccccgtcct tgtcaaatag ggagaaagcc
     3961 tctttaaatt ctgcgatctg ctcttcagtc agttggtccg gcaggttgta ctccagcttg
     4021 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     4081 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     4141 aagatggtgc gctcctggat gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     4201 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     4261 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     4321 taggtggcat cgccctcacc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     4381 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     4441 ctccctccgg taccgccctt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     4501 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttcga aagtttggac
     4561 tgcacgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     4621 tgctggtagt ggtaggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatgtggaag
     4681 ttcgccttga tgccgttctt ctgcttgtcg gccttgatat agacgttctc gagtgagctc
     4741 agccgaccta tagctctgac tgcgtgacct gtcttattcc acttacgacg tgatgagtcg
     4801 accatggtgg cgagatcctt atcgtcatcg tcgtacagat cccgacccat ttgctgtcca
     4861 ccagtcatgc tagccatacc atgatgatga tgatgatgag aaccaatctt ttggtcctca
     4921 tcattctttt caacctgttt ggttcttctc atacagcaca gcatggtggc gggtggatcc
     4981 ttgctagcag cttgaattcg actgcgctct caggcacgac acgactcctc cgctgcccac
     5041 cgcagactga ggcagcgctg agtcgccggc gccgcagcgc agatggtcgc gcccgtgccc
     5101 ccctatctcg cgcctcgcgt ggtgcggtcc ggctgggccg gcggcggcgc ggacgcgacc
     5161 aaggtggccg ggaaggggag tttgcggggg accggcgagt gacgtcagcg cgccttcagt
     5221 gctgaggcgg cggtggcgcg cgccgccagg cgggggcgaa ggcactgtcc gcggtgctga
     5281 agctggcagt gcgcacgcgc ctcgccgcat cctgtttccc ctccccctct ctgatagggg
     5341 atgcgcaatt tggggaatgg gggttgggtg cttgtccagt gggtcggggt cggtcgtcag
     5401 gtaggcaccc ccaccccgcc tcatcctggt cctaaaaccc acttgcactc atacgcaggg
     5461 ccctctgcag agatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     5521 attaactaca ag
//
