LOCUS       sequence-313160-        6616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ribosome-tagged GCaMP for soma localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  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 6616)
  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: pyc126m
FEATURES             Location/Qualifiers
     source          1..6616
                     /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..1830)
                     /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    2561..3149
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2614..2634)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3032..3043)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3181..3657
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3380..3399)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3697..3837
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3697..3826
                     /label=AAV2 ITR
     rep_origin      3912..4367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3999..4018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4209..4230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4384..4403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4503..4525
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4563..4581)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4649..4753
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4754..4822,4823..5614)
                     /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(4972..4991)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5785..6373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6274..6293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6435..6564
                     /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 tcaggccgga ctcagatctc aagctcaagc
     1861 ttcgagttca gcagcaaagt ctcacgcgac accctgtacg aggcggtgcg ggaagtcctg
     1921 cacgggaacc agcgcaagcg ccgcaagttt ctggagacgg tggagctgca gatcagcctg
     1981 aagaactacg accctcagaa ggacaaacgt ttctcgggca ccgtcaggct caagtccacc
     2041 ccacgcccca agttctcggt gtgcgttctg ggggaccagc agcactgtga tgaagccaag
     2101 gccgtggata tcccccacat ggacatcgag gcgctcaaga aacttaacaa aaacaagaag
     2161 ttggtcaaga agctggctaa gaagtacgat gcctttttgg cctctgagtc tctgattaag
     2221 cagatcccac gtatcctggg cccaggccta aacaaggctg gcaagttccc ctccctgctg
     2281 acacacaatg aaaacatggt ggccaaagtg gatgaggtga aatcgacaat caagttccag
     2341 atgaagaagg tgctgtgttt ggccgtcgct gttggccacg tgaagatgac cgatgatgag
     2401 ctagtctaca acattcatct ggctgtcaat ttcttggtgt ccttgcttaa gaaaaactgg
     2461 caaaacgtgc gggctctgta catcaagagc accatgggca agccccagcg tctgtattag
     2521 ggcgcgcccc tgcagggaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     2581 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2641 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2701 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2761 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2821 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2881 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2941 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3001 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3061 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3121 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     3181 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3241 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3301 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     3361 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     3421 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     3481 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     3541 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     3601 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     3661 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3721 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3781 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3841 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3901 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3961 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     4261 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4381 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4441 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4501 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4561 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4621 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4681 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4741 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4801 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4861 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4921 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4981 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5041 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5101 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5161 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5221 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5281 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5341 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5401 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5461 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5521 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5581 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5641 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5701 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5761 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5821 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5881 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5941 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6001 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6061 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6121 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6181 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6241 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6301 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6361 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6421 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     6481 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     6541 ccaactccat cactaggggt tcctgcggcc gcacgcgttt aattaagtgt ctagactgca
     6601 gagggccctg cgtatg
//
