LOCUS       sequence-358674-        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  JOURNAL   Unpublished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            "Unpublished"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 6f wpre  Pzac2.1
FEATURES             Location/Qualifiers
     source          1..6271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          402..534
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     517..536
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             548..925
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      938..947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(944..952,953..970,971..973,974..1006,1007..1009,
                     1010..1033,1034..1060,1061..1117,1118..1123,1124..1393,
                     1394..1411,1412..1846,1847..1852,1853..2296)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1040..1049
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1330..1351
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1457..1478)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1718..1737)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2359..2947
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2412..2432)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2830..2841)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3025..3146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3036..3055)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3090..3109
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3308..3437
                     /label=AAV2 ITR
     rep_origin      complement(3674..4262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3754..3773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4433..5224,5225..5293))
                     /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     5056..5075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5294..5398)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5424..5879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5561..5582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5773..5792
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5951..6080
                     /label=AAV2 ITR
ORIGIN
        1 atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg
       61 cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt
      121 cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc
      181 ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc
      241 gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg
      301 agagcgcagc tgtgctcctg ggcaccgcgc agtccgcccc cgcggctcct ggccagacca
      361 cccctaggac cccctgcccc aagtcgcagc cgttggatca ggtaagtatc aaggttacaa
      421 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
      481 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgatg
      541 atccggtatg ccagagccag cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc
      601 ggtgactaag gcgcagaaga aaggcggcaa gaagcgcaag cgcagccgca aggagagcta
      661 ttccatctat gtgtacaagg ttctgaagca ggtccaccct gacaccggca tttcgtccaa
      721 ggccatgggc atcatgaatt cgtttgtgaa cgacattttc gagcgcatcg caggtgaggc
      781 ttcccgcctg gcgcattaca acaagcgctc gaccatcacc tccagggaga tccagacggc
      841 cgtgcgcctg ctgctgcctg gggagttggc caagcacgcc gtgtccgagg gtactaaggc
      901 catcaccaag tacaccagcg ctaaggatcc accagtcgcc accatgggtt ctcatcatca
      961 tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga
     1021 cgatgacgat aaggatctcg ccaccatggt cgactcatca cgtcgtaagt ggaataagac
     1081 aggtcacgca gtcagagcta taggtcggct gagctcactc gagaacgtct atatcaaggc
     1141 cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg
     1201 cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1261 gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa
     1321 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1381 cgagctgtac aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac
     1441 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1501 gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1561 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     1621 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1681 cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1741 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1801 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacctgc cggaccaact
     1861 gactgaagag cagatcgcag aatttaaaga ggaattctcc ctatttgaca aggacgggga
     1921 tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac
     1981 agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacaatcga
     2041 cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa tacagggaca cggaagaaga
     2101 aattagagaa gcgttcggtg tgtttgataa ggatggcaat ggctacatca gtgcagcaga
     2161 gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat
     2221 gatcagggaa gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat
     2281 gatgacagcg aagtgaccgg atcactagtt cgcgatctag agaattctgc agatgatcca
     2341 ctcgagaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2401 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2461 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2521 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2581 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2641 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2701 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2761 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2821 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2881 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2941 tccccgcatc gataccgtcg acccgggcgg ccgctcgagc atgcatctag agtcgacccg
     3001 ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     3061 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3121 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3181 ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3241 tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     3301 aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3361 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     3421 agcgagcgag cgcgcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3481 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4081 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4381 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4501 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4621 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4861 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4921 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5041 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5101 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5281 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5401 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     5461 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5521 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5581 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5641 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5701 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5761 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5821 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     5881 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5941 attacgccag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
     6001 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     6061 ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta
     6121 gccatgctct aggaagatcc gagctccagt gtgctggaat tcgccctttt cagtatttaa
     6181 attgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
     6241 tcccctatca gagaggggga ggggaaacag g
//
