LOCUS       sequence-210566-        5986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For enriched expression of GCaMP6m in axons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5986)
  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 5986)
  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-GCaMP6m
FEATURES             Location/Qualifiers
     source          1..5986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     promoter        470..917
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1012..1017,1018..1035,1036..1038,1039..1071,1072..1074,
                     1075..1098,1099..1125,1126..1182,1183..1188,1189..1458,
                     1459..1476,1477..1911,1912..1917,1918..2361)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6m
                     /translation="GSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNKT
                     GHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSL
                     GQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGNG
                     YISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1105..1114
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1395..1416
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1522..1543)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1783..1802)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2374..2962
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2427..2447)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2845..2856)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2965..2981)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2966..2982)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3011..3132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3022..3041)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3076..3095
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3294..3423
                     /label=AAV2 ITR
     primer_bind     complement(3442..3459)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3442..3458)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3451..3473)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3600..4055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3687..3706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3897..3918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4081..4185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4186..4254,4255..5046)
                     /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(4404..4423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5217..5805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5706..5725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg
      481 gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac
      541 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      601 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      661 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      721 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      781 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      841 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
      901 cgtgcctgag agcgcagtcg aattcaagct gctagcaagg atcccgccac catgctgtgc
      961 tgtatgagaa gaaccaaaca ggttgaaaag aatgatgagg accaaaagat tggttctcat
     1021 catcatcatc atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg
     1081 tacgacgatg acgataagga tctcgccacc atggtcgact catcacgtcg taagtggaat
     1141 aagacaggtc acgcagtcag agctataggt cggctgagct cactcgagaa cgtctatatc
     1201 aaggccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca caacatcgag
     1261 gacggcggcg tgcagctcgc ctaccactac cagcagaaca cccccatcgg cgacggcccc
     1321 gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca aactttcgaa agaccccaac
     1381 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1441 atggacgagc tgtacaaggg cggtaccgga gggagcatgg tgagcaaggg cgaggagctg
     1501 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1561 agcgtgtccg gcgagggtga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1621 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1681 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1741 atgcccgaag gctacatcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1801 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1861 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa cctgccggac
     1921 caactgactg aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac
     1981 ggggatggga caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac
     2041 cccacagaag cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca
     2101 atcgacttcc ctgagttcct gacaatgatg gcaagaaaag ggagctacag ggacacggaa
     2161 gaagaaatta gagaagcgtt cggtgtgttt gataaggatg gcaatggcta catcagtgca
     2221 gcagagcttc gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat
     2281 gaaatgatca gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta
     2341 caaatgatga cagcgaagtg aaagcttatc gataatcaac ctctggatta caaaatttgt
     2401 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2461 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2521 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2581 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2641 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2701 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2761 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2821 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2881 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2941 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gacccgggcg gccgcttcga
     3001 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3061 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3121 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     3181 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt
     3241 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     3301 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3361 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3421 cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     3481 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     3541 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     3601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3661 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3781 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3901 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3961 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4021 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     4081 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4141 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4201 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4261 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4321 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4381 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4441 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4501 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4561 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4621 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4681 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4741 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4801 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4861 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4921 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4981 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5041 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5101 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5161 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5221 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5281 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5341 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5401 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5461 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5521 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5581 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5641 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5701 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5761 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5821 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5881 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5941 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
