LOCUS       sequence-381764-        6560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ChemoG-CaM in primary neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6560)
  AUTHORS   Hellweg L, Edenhofer A, Barck L, Huppertz MC, Frei MS, Tarnawski M, 
            Bergner A, Koch B, Johnsson K, Hiblot J
  TITLE     A general method for the development of multicolor biosensors with 
            large dynamic ranges.
  JOURNAL   Nat Chem Biol. 2023 Jun 8. doi: 10.1038/s41589-023-01350-1.
  PUBMED    37291200
REFERENCE   2  (bases 1 to 6560)
  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: "Nat Chem 
            Biol. 2023 Jun 8. doi: 10.1038/s41589-023-01350-1."
COMMENT     Sequence Label: pAAV2-hSyn1-NES-ChemoG-CaM
FEATURES             Location/Qualifiers
     source          1..6560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..519)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             709..1149
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     misc_feature    2239..2827
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2292..2312)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2710..2721)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2830..2846)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2831..2847)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2876..2997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2887..2906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2941..2960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3159..3288
                     /label=AAV2 ITR
     rep_origin      complement(3525..4113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3605..3624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4284..5075,5076..5144))
                     /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     4907..4926
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5145..5249)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5275..5730)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5412..5433)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5624..5643
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5802..5931
                     /label=AAV2 ITR
     promoter        6013..6460
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actggtggca ccctgccgga ccaactgact
      721 gaagagcaga tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg
      781 acaataacaa ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa
      841 gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc
      901 cctgagttcc tgacaatgat ggcaagaaaa atgaaagaca cagacagtga agaagaaatt
      961 agagaagcgt tccgtgtgtt tgataaggat ggcaatggct acatcagtgc agcagagctt
     1021 cgccacgtga tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc
     1081 agggaagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt agtgatgatg
     1141 acagcgaagg aattccctcc tcctccgcca ccgccaccac cgccgccgcc gccaccacca
     1201 cctcctccgc ctcctccacc accaccgcca ccgcctccac cacctggcgg cagcagggtc
     1261 gactcatcac gtcgtaagtt caataagaca ggtaaggcac tgagagctat aggtcggctg
     1321 agctcactcg agtctggcgg tatcggtact ggctttccat tcgaccccca ttatgtggaa
     1381 gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg
     1441 ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc gcatgttgca
     1501 ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga caaaccagac
     1561 ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga agccctgggt
     1621 ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt ccactgggcc
     1681 aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg ccctatcccg
     1741 acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg caccaccgac
     1801 gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct gccgatgggt
     1861 gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt cctgaatcct
     1921 gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg tgagccagcg
     1981 aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc tgtcccgaag
     2041 ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc
     2101 aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtgaaaatct gctgcaagaa
     2161 gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct ggagatttcc
     2221 ggttgaaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2281 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2341 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2401 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2461 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2521 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2581 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2641 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2701 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2761 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2821 tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga
     2881 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2941 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3001 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3061 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     3121 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     3181 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3241 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga
     3301 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3361 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3421 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3481 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3541 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3601 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3661 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3721 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3781 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3841 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3901 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3961 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4021 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4081 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4141 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4201 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4261 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4321 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4381 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4441 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4501 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4561 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4621 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4681 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4741 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4801 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4861 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4921 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4981 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5041 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5101 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5161 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5221 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     5281 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5341 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5401 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5461 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     5521 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5581 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5641 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5701 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     5761 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca
     5821 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     5881 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
     5941 tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc tctgcagagg
     6001 gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta
     6061 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
     6121 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactaccagc
     6181 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
     6241 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
     6301 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
     6361 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
     6421 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgaattcaa gctgctagca
     6481 aggatccacc cgccaccatg ctgcagaacg agcttgctct taagttggct ggacttgata
     6541 ttaacaagac tggaggttct
//
