LOCUS       sequence-355293-        6271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  calcium sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6271)
  TITLE     1.	AAV  Synapsin  Intron H2B Gcamp 6f wpre  Pzac2.1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6271)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AAV Synapsin Intron H2B Gcamp 6s wpre Pzac2.1
FEATURES             Location/Qualifiers
     source          1..6271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     polyA_signal    292..413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(329..348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     383..402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    491..1079
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             597..608
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1006..1026
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1142..1585,1586..1591,1592..2026,2027..2044,
                     2045..2314,2315..2320,2321..2377,2378..2404,2405..2428,
                     2429..2431,2432..2464,2465..2467,2468..2485,2486..2494))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1701..1720
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1960..1981
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2087..2108)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2389..2398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2491..2500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2513..2890)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2902..2921)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2904..3036
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   3629..3758
                     /label=AAV2 ITR
     rep_origin      3830..4285
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3917..3936)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4127..4148
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4311..4415
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4416..4484,4485..5276)
                     /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(4634..4653)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5447..6035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5936..5955
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcc ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc
      241 ccacatctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
      301 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
      361 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
      421 tgctcgaagc ggccgcccgg gtcgactcta gatgcatgct cgagcggccg cccgggtcga
      481 cggtatcgat gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga
      541 agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg
      601 agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg
      661 agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac
      721 agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata
      781 gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca
      841 gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac
      901 aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata
      961 cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga
     1021 gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta
     1081 tcgataagct tctcgagtgg atcatctgca gaattctcta gatcgcgaac tagtgatccg
     1141 gtcacttcgc tgtcatcatt tgtacaaact cttcgtagtt tacctgacca tccccatcga
     1201 tgtctgcttc cctgatcatt tcatcaacct cttcatctgt taacttctct ccaaggtttg
     1261 tcatcacgtg gcgaagctct gctgcactga tgtagccatt gccatcctta tcaaacacac
     1321 cgaacgcttc tctaatttct tcttccgtgt ccctgtattt catttttctt gccatcattg
     1381 tcaggaactc agggaagtcg attgtgccgt caccgtcggc atctacttca ttgatcatgt
     1441 cctgcagctc tgcttctgtg gggttctgcc ccagagaccg catcaccgtc cccagctcct
     1501 tggttgttat tgtcccatcc ccgtccttgt caaataggga gaaagcctct ttaaattctg
     1561 cgatctgctc ttcagtcagt tggtccggca ggttgtactc cagcttgtgc cccaggatgt
     1621 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     1681 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     1741 cctggatgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     1801 ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca
     1861 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     1921 cctcaccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     1981 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatgctc cctccggtac
     2041 cgcccttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     2101 ggaccatgtg atcgcgcttc tcgttggggt ctttcgaaag tttggactgc acgctcaggt
     2161 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     2221 aggcgagctg cacgccgccg tcctcgatgt tgtggcggat gtggaagttc gccttgatgc
     2281 cgttcttctg cttgtcggcc ttgatataga cgttctcgag tgagctcagc cgacctatag
     2341 ctctgactgc gtgacctgtc ttattccact tacgacgtga tgagtcgacc atggtggcga
     2401 gatccttatc gtcatcgtcg tacagatccc gacccatttg ctgtccacca gtcatgctag
     2461 ccataccatg atgatgatga tgatgagaac ccatggtggc gactggtgga tccttagcgc
     2521 tggtgtactt ggtgatggcc ttagtaccct cggacacggc gtgcttggcc aactccccag
     2581 gcagcagcag gcgcacggcc gtctggatct ccctggaggt gatggtcgag cgcttgttgt
     2641 aatgcgccag gcgggaagcc tcacctgcga tgcgctcgaa aatgtcgttc acaaacgaat
     2701 tcatgatgcc catggccttg gacgaaatgc cggtgtcagg gtggacctgc ttcagaacct
     2761 tgtacacata gatggaatag ctctccttgc ggctgcgctt gcgcttcttg ccgcctttct
     2821 tctgcgcctt agtcaccgcc ttcttggagc cctttttcgg ggcgggagca gacttcgctg
     2881 gctctggcat accggatcat cacctgtgga gagaaaggca aagtggatgt cagtaagacc
     2941 aataggtgcc tatcagaaac gcaagagtct tctctgtctc gacaagccca gtttctattg
     3001 gtctccttaa acctgtcttg taaccttgat acttacctga tccaacggct gcgacttggg
     3061 gcagggggtc ctaggggtgg tctggccagg agccgcgggg gcggactgcg cggtgcccag
     3121 gagcacagct gcgctctcag gcacgacacg actcctccgc tgcccaccgc agactgaggc
     3181 agcgctgagt cgccggcgcc gcagcgcaga tggtcgcgcc cgtgcccccc tatctcgcgc
     3241 ctcgcgtggt gcggtccggc tgggccggcg gcggcgcgga cgcgaccaag gtggccggga
     3301 aggggagttt gcgggggacc ggcgagtgac gtcagcgcgc cttcagtgct gaggcggcgg
     3361 tggcgcgcgc cgccaggcgg gggcgaaggc actgtccgcg gtgctgaagc tggcagtgcg
     3421 cacgcgcctc gccgcatcct gtttcccctc cccctctctg ataggggatg cgcaatttgg
     3481 ggaatggggg ttgggtgctt gtccagtggg tcggggtcgg tcgtcaattt aaatactgaa
     3541 aagggcgaat tccagcacac tggagctcgg atcttcctag agcatggcta cgtagataag
     3601 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     3661 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3721 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc
     3781 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     3841 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3901 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3961 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4021 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4081 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4141 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4201 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4261 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4321 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4381 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4441 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4501 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4561 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4621 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4681 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4741 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4801 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4861 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4921 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4981 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5041 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5101 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5161 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5221 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5281 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5341 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5401 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5461 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5521 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5581 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5641 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5701 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5761 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5821 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5881 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5941 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6001 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6061 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6121 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6181 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6241 tccccgcgcg ttggccgatt cattaatgca g
//
