LOCUS       sequence-313230-        9277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pan-neuronal nuclear-localized GCaMP expression in C. elegans .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9277)
  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 9277)
  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: pHJ4
FEATURES             Location/Qualifiers
     source          1..9277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4397..4417
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4418..4426,4427..4444,4445..4447,4448..4480,4481..4483,
                     4484..4507,4508..4534,4535..4591,4592..4597,4598..4867,
                     4868..4885,4886..5320,5321..5326,5327..5770)
                     /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      4514..4523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4804..4825
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4931..4952)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5192..5211)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(6695..6717)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6817..6836
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6918..6936)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7004..7108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7109..7177,7178..7969)
                     /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(7327..7346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8140..8728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8629..8648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8882..8899
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9016..9037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9052..9057,9058..9075,9076..9082)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9090..9106
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9095..9117
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9114..9130
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9114..9130
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcgagttttg actggctttc aaaaactgag tgattactaa ttatttttga aaaaattaac
       61 agttgtgata tgatatttag acatttttgc aatatataat ttgtttgaaa aaaaaacatt
      121 ttttaatggt cttcgaagct ataatagttt ttcgtaaaat tttaaataaa acagctgaca
      181 gatatccaca ttcaataatt gaaagtatgc agttctcttt ttttttgtca aatagaaaaa
      241 aagtaacaga tggtcagata taatctaggt taatctgact ctggtcatcg atgaactcat
      301 tttcatcata atcattcttc tttcaaatgg tctgttcgga attcgaatat tcaataaatg
      361 gatatcttat tagatgctcc ggatttattc caccaaactt ccaagacgta agttcaattt
      421 aaacaaaatt actatagctt gaaaaaatga gaaaaaacga gcagataaaa aaataacaga
      481 aaaaatccat aaaaatgacg agggttccgt gcgaaaatcg tcattgtttt tggcttgcat
      541 cacacttttc cgcgagtcag ccacactgtt ttatcgatta ttatttttct gatttcagaa
      601 accggaaact tacaactgag atgctttcat tttgattttt ttaaatttag ttttgatttc
      661 agcgatttgt ttgaatataa ttatgcatga gttggctttt gcatttttag ataaataagt
      721 tcacatgctc ggacaaattg tgtggtttaa tctgtgaaag ttttgaatct tctgtttgaa
      781 aaaaaaaaca tttttaattt gtatatacga gtgaattggt taggccctgg ttacgttttc
      841 cagcgaattc attcaaattt cttatttttt ctattttttc aattaaataa tttagttctc
      901 tatcacatgt gtatgatcaa aatttcattc ctaatcgtaa gcaaatttag gctcatccgt
      961 agtataattt tgttatgaag ggcttcacag taaaagctca aactccaaca tttagcaact
     1021 agcaaccaac aaaaaaccaa agaaatgtca gttttgaaaa gtcgttattt ctttgtcttc
     1081 tcacaattca cacatcatct tctcgttttc ctccgttttt ttgtcgtgct tccttctttt
     1141 ctttacacat ttttctaacc tttttttaag tctatcctct cctaatcctt tgtactctct
     1201 aatcagtttt gctggttgtg ctgcacaatt tttatcagat ttatgcgcga aacaaaagcc
     1261 tctcgcggag agctccgcca aatccggtct tcttttttcc ttgcctatcg gatttcacaa
     1321 acattcagtt tgtcacttga cgatgtgttg gaggcattac tactaggaag tgttattcat
     1381 attttttatt cgtttctttt tggtgactaa acgaaatatt ccgaacgttt tcctgatttc
     1441 aggcggcgga aacgaagaag accacgaata atcaacaggc tgccattgca tccgccagaa
     1501 gtcgagtgag ttctttgaaa atctttatta gagttatttg tgacaatttg ccactttttt
     1561 tttaatgttt aatgaatctg taacgaaatt aataccaagg gtgtccaaca gaactatctg
     1621 agcggttatt taatttttag atagtatatt tcttccgatc gtaacataca tattttcaag
     1681 catagtactg tgatttaagt tacagttttc caatcagtct tgattttaat aaaaactttt
     1741 caactgcaaa aatgcctctg aagtattttt ctatattttt atagttgaat aattttttga
     1801 ttggcacaaa gaccgccgag atatgagccg ttaaagtaaa gctgcatgga ttcaatatgg
     1861 tttgtatttg attggagtca cttacatgga aatatttgta tttaaatctc atattttaaa
     1921 acaagacaaa caccaaatta aatttttatt aatctgaaca tttttcaatc gatttggtaa
     1981 acaaattatt agatgtttcc ctcaaaaatt gtaacaattc ctaaattgtg gcacttataa
     2041 taacatcgtc ctgaattctc aaaaaaaacc gaatgctaag agagatttcg tttaactggg
     2101 acctgtaact gaattttcaa atattggcag tttttgataa actgggaggg aaatcaaaaa
     2161 ccaaaactac agtacccctc gtgttataat aaattgtcat tcactccttt tttgttccca
     2221 acatttgctc ccatctcctg acgagaaaga acaattttat gagcacggga acccgtgggc
     2281 gtccatccgt tcctcctgtg caatttcagc ccccttcctc tacattctct gtgttcccgt
     2341 ttttatcgtt ttatgggtat cgggaacgga cagatcgctt ccatttgctt ctattccgtc
     2401 ctcacaagac gtcccactaa ctgattttct tctgatttct tttcactgaa aatgggaaag
     2461 aaagaaaaag acgatgaatc gacgacttct tctcaggaga aggtgaagaa gaaggataag
     2521 gagaaagttc atctaaggct gaaagataaa gtaaaaaaac ttagaaaatc taagtcgatt
     2581 gcagtatctt ctccagctga ttatatgact tgtcagtgtg aaccatgcaa aaaagtagct
     2641 ctcgaggtaa taatttttaa actcaatacg tagttaagtt tgaactcctt ttaatttact
     2701 tatcatctaa tttaccgtca caatagtact tccattagca attattttaa aattacatgc
     2761 atagcttcaa tgttcgtttt tttgcgttct catatcgtat gttgtctgcc tctcgaatac
     2821 tgactgtccg actagaatgt cccggtagga ttactgtacg gtcactggga ataaactgtt
     2881 tctgaaatca atctgaaatt ccgtagatct tcttggttca atttcttatt cctcaatctt
     2941 tggacaacgt caccttcaga gtcgtcgttg agtatcccat cattaactag tcaatctttc
     3001 tctctttttc atttccccca acttttctgc atattttttg acgacgacga cctcgacggc
     3061 aacgagaata catttatcta ttttttagta tagcatatat acattttcta cttttctcta
     3121 acccatttca gacgtataga ttttggtttt cgaatgcaac tttgagattt ttcaaagatc
     3181 ttcagatggg agcagtggac tgtctatttg atattcttaa aatgtactct ataaaagatg
     3241 aaactgtaat gaccaattcc ctacataaat atttttaact aattggtcgg atatttgggg
     3301 atcagagaag tacttgtttt tgcgatatta ataatgatat atttttctag taaaaaaatg
     3361 ttaagcgatt tgtgaataat taagttggct tgaaaaagat tttttgatag ctatgtattt
     3421 tgttaattta aatcagattt taaaacttta ttattgacta tttatatata ttatatatta
     3481 tatattttca tggtttaatc aaaatctatt aaattatttc taagatctct aaatatcgaa
     3541 ttctctaaaa agcttcatca acaaaagcat ttcaattatc taacacaatg agaataattc
     3601 aagaactgaa atagaaagaa atctagaaaa aaaagagagt gaggtgccct attaaatcgg
     3661 tgctctcctc tctaacagaa gcgcctacac gcacccaaga gagctctttt tctttctttt
     3721 ttcggagcac acatacacgc atacactcct cagaagtgta tagagaaaat gggagaagaa
     3781 gcgaaagaag acgaagaagc ggttgtgatg tatgcatcgg gcgtttctat ccctctccgt
     3841 gagcaacgag ctagtcaacc catcgatcca tttttgtgag cacacacaga gagagactca
     3901 aacacactca cagcttcaca ctgagctcca agagctctgt agagagaagc cagagtatgc
     3961 tttcctcgat tttcctaacg tcttcttctt ctgctttttt ctttttccca acgagttgta
     4021 gtccactaac ttcaattttt cctccagaga aagagtttta gagagacagc tgataaatac
     4081 actagagaaa taaaaagaat gataaaaatt tgctatttta atcagttccc tctcgtttct
     4141 ctaagttttg ttgttgggtt gggaaattaa aattggaaaa tggaaataac gtttttttct
     4201 gtctaatctt gaattctcca gacatttctt atttcctttt ttgttgcaag tttttcttcc
     4261 ttctctaatt tttaatcaaa gatatctcgc tatgaacttt gaatttttga tattttgatt
     4321 ctataaatac ataaagtaat ctaaatgctc ttttaaaata aatctacagt agccctattt
     4381 tcagggatcc atggctccaa agaagaagcg taaggtaatg ggttctcatc atcatcatca
     4441 tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga
     4501 cgataaggat ctcgccacca tggtcgactc atcacgtcgt aagtggaata agacaggtca
     4561 cgcagtcaga gctataggtc ggctgagctc actcgagaac gtctatatca aggccgacaa
     4621 gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt
     4681 gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4741 cgacaaccac tacctgagcg tgcagtccaa actttcgaaa gaccccaacg agaagcgcga
     4801 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     4861 gtacaagggc ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt
     4921 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4981 cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     5041 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     5101 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     5161 ctacatccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     5221 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     5281 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga
     5341 agagcagatc gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac
     5401 aataacaacc aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc
     5461 agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc
     5521 tgagttcctg acaatgatgg caagaaaagg gagctacagg gacacggaag aagaaattag
     5581 agaagcgttc ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg
     5641 ccacgtgatg acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag
     5701 ggaagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac
     5761 agcgaagtga gggcgccatg agccgtagac gaaaagcgaa tccgacaaaa ctgagtgaaa
     5821 acgcgaagaa gcttgccaag gaagttgaaa attaaggtac catggtattg atatctgagc
     5881 tccgcatcgg ccgctgtcat cagatcgcca tctcgcgccc gtgcctctga cttctaagtc
     5941 caattactct tcaacatccc tacatgctct ttctccctgt gctcccaccc cctatttttg
     6001 ttattatcaa aaaacttctc ttaatttctt tgttttttag cttcttttaa gtcacctcta
     6061 acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat
     6121 tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta
     6181 cacttcttat gttttttact tctgataaat ttttttgaaa catcatagaa aaaaccgcac
     6241 acaaaatacc ttatcatatg ttacgtttca gtttatgacc gcaattttta tttcttcgca
     6301 cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg gagtattttt
     6361 ggaatttttc aatcaagtga aagtttatga aattaatttt cctgcttttg ctttttgggg
     6421 tttcccctat tgtttgtcaa gatttcgagg acggcgtttt tcttgctaaa atcacaagta
     6481 ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct cagtggaagg
     6541 tgagtagaag ttgataattt gaaagtggag tagtgtctat ggggtttttg ccttaaatga
     6601 cagaatacat tcccaatata ccaaacataa ctgtttctac tagtcggccg tacgggccct
     6661 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6721 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6781 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6841 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6901 ggcggcctta attaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6961 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7021 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7081 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7141 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7201 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7261 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7321 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7381 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7441 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7501 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7561 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7621 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7681 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7741 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7801 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7861 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7921 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7981 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8041 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     8101 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     8161 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     8221 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     8281 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     8341 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     8401 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     8461 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     8521 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     8581 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     8641 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8701 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8761 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     8821 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     8881 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     8941 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     9001 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     9061 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     9121 cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat
     9181 tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct
     9241 aacaacttgg aaatgaaata agcttgcatg cctgcag
//
