LOCUS       sequence-313163-        5929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  soluble GCaMP expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5929)
  AUTHORS   Chen Y, Jang H, Spratt PWE, Kosar S, Taylor DE, Essner RA, Bai L, 
            Leib DE, Kuo TW, Lin YC, Patel M, Subkhangulova A, Kato S, Feinberg 
            EH, Bender KJ, Knight ZA, Garrison JL
  TITLE     Soma-Targeted Imaging of Neural Circuits by Ribosome Tethering.
  JOURNAL   Neuron. 2020 Jun 5. pii: S0896-6273(20)30351-2. doi: 
            10.1016/j.neuron.2020.05.005.
  PUBMED    32574560
REFERENCE   2  (bases 1 to 5929)
  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: "Neuron. 
            2020 Jun 5. pii: S0896-6273(20)30351-2. doi: 
            10.1016/j.neuron.2020.05.005."
COMMENT     Sequence Label: pyc128m
FEATURES             Location/Qualifiers
     source          1..5929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      475..484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(481..489,490..507,508..510,511..543,544..546,547..570,
                     571..597,598..654,655..660,661..930,931..948,949..1383,
                     1384..1389,1390..1833)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6m
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     867..888
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(994..1015)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1255..1274)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1874..2462
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1927..1947)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2345..2356)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2494..2970
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2693..2712)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3010..3150
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3010..3139
                     /label=AAV2 ITR
     rep_origin      3225..3680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3312..3331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3522..3543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3697..3716)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3816..3838
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3876..3894)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3962..4066
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4067..4135,4136..4927)
                     /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(4285..4304)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5098..5686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5587..5606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5748..5877
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg ctcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaaccgg ctagaggatc cggcgccacc
      481 atgggttctc atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg
      541 ggtcgggatc tgtacgacga tgacgataag gatctcgcca ccatggtcga ctcatcacgt
      601 cgtaagtgga ataagacagg tcacgcagtc agagctatag gtcggctgag ctcactcgag
      661 aacgtctata tcaaggccga caagcagaag aacggcatca aggcgaactt caagatccgc
      721 cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc
      781 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc caaactttcg
      841 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      901 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag
      961 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1021 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     1081 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1141 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1201 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac
     1261 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1321 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1381 aacctgccgg accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta
     1441 tttgacaagg acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct
     1501 ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac
     1561 ggtgacggca caatcgactt ccctgagttc ctgacaatga tggcaagaaa agggagctac
     1621 agggacacgg aagaagaaat tagagaagcg ttcggtgtgt ttgataagga tggcaatggc
     1681 tacatcagtg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat
     1741 gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac
     1801 gaagagtttg tacaaatgat gacagcgaag tgaggcgcgc ccctgcaggg aattcgatat
     1861 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1921 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     1981 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2041 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2101 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2161 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2221 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     2281 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2341 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2401 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2461 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     2521 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     2581 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     2641 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     2701 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     2761 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     2821 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     2881 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     2941 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     3001 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3061 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     3121 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     3181 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     3241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3301 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3361 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3421 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3541 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     3601 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3661 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     3721 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     3781 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     3841 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     3901 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3961 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4021 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4081 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4141 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4201 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4261 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4321 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4381 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4441 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4501 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4561 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4621 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4681 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4741 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4801 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4861 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4921 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4981 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5041 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5101 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5161 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5221 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5281 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5341 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5401 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5461 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5521 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5581 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5641 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5701 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
     5761 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
     5821 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     5881 gccgcacgcg tttaattaag tgtctagact gcagagggcc ctgcgtatg
//
