LOCUS       sequence-313237-        8139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ribosome-tagged GCaMP (soma localized) expression pan-neuronally C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8139)
  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 8139)
  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: pAS18
FEATURES             Location/Qualifiers
     source          1..8139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(2794..2802,2803..2820,2821..2823,2824..2856,2857..2859,
                     2860..2883,2884..2910,2911..2967,2968..2973,2974..3243,
                     3244..3261,3262..3696,3697..3702,3703..4143)
                     /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      2890..2899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3180..3201
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3307..3328)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3568..3587)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5656..5678)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5778..5797
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5879..5897)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5965..6069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6070..6138,6139..6930)
                     /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(6288..6307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7101..7689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7590..7609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7843..7860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7977..7998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8013..8018,8019..8036,8037..8043)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8051..8067
                     /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     8056..8078
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8075..8091
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8075..8091
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggatccatag gtatgcctgt cattctctat ttttaaacgt tcctgcagca gtagcacaac
       61 agagccagca gttttcctct tcttctccat cttcttctgc ttcttcctct cttcccaaaa
      121 aaaaaaaaaa aaaaaaaagt gagggggctc actgcccccc cccccctccc ccccccctct
      181 tctccttgaa gctcatccag acgtcgctgt cctcctcctc ctcgtccccg gggcgctgtg
      241 tgtgtgtgtg tgtgctccgt gagcatgtag cagcttcagc ccatttaaag gcgcacactc
      301 tctgcatctc tgcgtgtctc gacactcgtt ttttctcttc ttttctgctg cacacacaca
      361 cacacacaca cacacgctca tgccagagca cttcttgctc cggggggctc cgcgcggagc
      421 ccgagaggac ctcctcctcc tttttcttct tcgcttttta gttcttcttg cctcggtctc
      481 gccccttgtt ttgctctcca tgtagtatat acatctgggt gtgtgtgtgt gtgtgcacct
      541 ttggagcatt ttttgaacct ttccagaact ttggagcaac attttcagct tgttcagtgc
      601 actttttagg ctgaaaatcc tatttttagg ttttcgaaag tttttttgga gcatttttcg
      661 tgcaatttag gtgcaaaatt aaaaaaaagg gttttgtagt gaattaaaat ttttttgaaa
      721 ctttcgcctt tttaaaaaaa tgtttctaat atctttaaaa attggaatat tttggaaaac
      781 aatttgtttt caattggcaa tttattaatt ctgagcttct gagcgtaaat ttttttttat
      841 tttggagcaa ttctgagatg tcttagtgca tttttcgggc atacataaaa aaaaatttat
      901 cttaataata aaaaccttaa atttaggctt caccgtctgg aaattcaatt ttttcagctg
      961 aaaaactttc aaaaaaaaaa aaattaaatt ttcacattta ttttctctgg aattttggag
     1021 caatttcaaa gctgtcttag tgcattttta gtacacaaat attttttcat ttttcttaaa
     1081 ttttaataat ctaagcttaa ataaaggctt caccgtctga aaatgcaact tttttcagat
     1141 gaaatattaa aaaaaaaacc tcttttatta attttgtgct ccaaactttc ccacggtacg
     1201 caaatcatgt aatatgtact tttaaacatt atttttccaa tcaaaaatat ccaacagctt
     1261 ttccttacaa tttgaaaatt tttattaaaa tttcaaaaaa attttttttt tttttaattt
     1321 cgaatttttt taaacttgtt gtaacttttt tccctgtcgg cgccaattaa caaaatacgt
     1381 agaattttgc gcttattgtt ttgttagtta acaaattttt gacaactttc ttatcagctg
     1441 agatttaggt gtttcagttc gcaatctgcg tctctccttg ctctcctcta ttttaaaacc
     1501 cccataactt tgttcttgtg agagttatca aaaaattgtc aactaacaaa atatagagca
     1561 caaaaatatg cacattttgt atgttgacaa ttttttgata ccttttttag ttttcgagat
     1621 atcttagttt tagtttagct gtaccttttt ctggcaaaaa acttccaaat ttcggcaatt
     1681 tttcggcaaa tttcccggca aatttcgcca agtacatcgg gctctcagac aagtggtctg
     1741 tgagtggaag tcgataaaag tagtttacgt gctctccgtc cccccgcttc ttctccgtct
     1801 ctgcttctat ttctccattg actagtccca ctacttttta cggggcggag ggctctcttg
     1861 cactccagct tcttctcttt ttgtgctctt ttttgcccgt atgtgggtgc tcatttattt
     1921 ttctttggtt ttaaaataaa ttcggtttat taggctactc gtaaaaatga agaaaaaact
     1981 tcaaattcaa acgacattaa aaaaaaattt aataaaaatt ttttgaattg ctcaaaagcc
     2041 ccacctatat atatatatag ctccgcctac ttctcgcaca gaccccgccc actttttgac
     2101 tccgccccct tattggcaaa agttcttgaa aaaaataaaa ataaaaattc aaattgaaac
     2161 aatttttttt taatgaagta gtttttttct aaattgatga aaatttctaa acagtttaca
     2221 agcaccgccc actttttgac tccgcccctt tgttggaaaa attcttgtaa aaaaaaataa
     2281 aaaataaaag cttctctttc aaacgatttt tagttggtct ttgtatgaaa agtttttttc
     2341 ctaaaataga aatgtctgaa taatttgcaa gctccgccct tttcttaccc agactccgcc
     2401 cctttattga ctaatttttt tttgtccatt tacctgaagt tcaaaaaaaa aatttacagt
     2461 ttcatagagt aataattttt caaaaatctc tttttccaat gtttaatgtg tcattataaa
     2521 cccgcccagt tttaagctcc gcccatttag gccccgccca tttatgaccc aaatattttt
     2581 ttctctattc acccgtggat ttccctccaa caagttaccc caaaaataaa aataaaaatt
     2641 cctgcaggcg cccaccggga tccggcgcgc ctctagagga tccccgggat tggccaaagg
     2701 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
     2761 ggctagcgtc gacggtacca tggtattgat atcatgggtt ctcatcatca tcatcatcat
     2821 ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat
     2881 aaggatctcg ccaccatggt cgactcatca cgtcgtaagt ggaataagac aggtcacgca
     2941 gtcagagcta taggtcggct gagctcactc gagaacgtct atatcaaggc cgacaagcag
     3001 aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     3061 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3121 aaccactacc tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac
     3181 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3241 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg
     3301 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3361 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3421 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     3481 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3541 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3601 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3661 gacggcaaca tcctggggca caagctggag tacaacctgc cggaccaact gactgaagag
     3721 cagatcgcag aatttaaaga ggctttctcc ctatttgaca aggacgggga tgggacaata
     3781 acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag
     3841 ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag
     3901 ttcctgacaa tgatggcaag aaaagggagc tacagggaca cggaagaaga aattagagaa
     3961 gcgttcggtg tgtttgataa ggatggcaat ggctacatca gtgcagcaga gcttcgccac
     4021 gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa
     4081 gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg
     4141 aagtcaggcc ggactcagat ctcaagctca agcttcgagt tctcgaaggt ttcccgcgag
     4201 tctcttaatg aggctatcgc tgaggtcctt aagggatcct cggaaaagcc acgcaagttc
     4261 cgtgagacta tcgagctcca gatcggtctc aagaactacg acccacagaa ggacaagcgt
     4321 ttcagcggat ccatcagact gaagcacatt ccacgcccaa acatgaaggt gtgcgtcttc
     4381 ggagaccagc accatcttga cgaggctgcc gccggagaca ttccatcgat gagcgccgat
     4441 gacttgaaga agcttaacaa gcagaagaag ctcatcaaga agctcgccaa gagctacgat
     4501 gctttcatcg cttccgaatc cttgatcaag cagatcccac gtatcctcgg tccaggactc
     4561 aacaaggccg gaaaattccc ttccgtcgtt acccacggag aatctctcca aagcaagagt
     4621 gacgagatcc gcgccaccgt caagttccag atgaagaagg ttctctgcct ctccgtcgcc
     4681 gtcggtcacg ttggactcac ccaggaggag cttgtctcca acatctccct ctccatcaac
     4741 ttcctcgtct cgctcttgaa gaagaactgg cagaacgtga gatcgttgaa catcaagtcc
     4801 accatgggaa agccacaaag agtttattaa gatatctgag ctccgcatcg gccgctgtca
     4861 tcagatcgcc atctcgcgcc cgtgcctctg acttctaagt ccaattactc ttcaacatcc
     4921 ctacatgctc tttctccctg tgctcccacc ccctattttt gttattatca aaaaacttct
     4981 cttaatttct ttgtttttta gcttctttta agtcacctct aacaatgaaa ttgtgtagat
     5041 tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc tccccccatt
     5101 aataataatt ctatcccaaa atctacacaa tgttctgtgt acacttctta tgttttttac
     5161 ttctgataaa tttttttgaa acatcataga aaaaaccgca cacaaaatac cttatcatat
     5221 gttacgtttc agtttatgac cgcaattttt atttcttcgc acgtctgggc ctctcatgac
     5281 gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt tggaattttt caatcaagtg
     5341 aaagtttatg aaattaattt tcctgctttt gctttttggg gtttccccta ttgtttgtca
     5401 agatttcgag gacggcgttt ttcttgctaa aatcacaagt attgatgagc acgatgcaag
     5461 aaagatcgga agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa gttgataatt
     5521 tgaaagtgga gtagtgtcta tggggttttt gccttaaatg acagaataca ttcccaatat
     5581 accaaacata actgtttcta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg
     5641 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5701 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5761 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5821 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt aattaagggc
     5881 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5941 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6001 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6061 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6121 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6181 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6241 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6301 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6361 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6421 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6481 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6541 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6601 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6661 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6721 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6781 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6841 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6901 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6961 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7021 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7081 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7741 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7801 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7861 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7921 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7981 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8041 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8101 gccaagcttg catgcgcggc cgcactgact gggccggcc
//
