LOCUS       sequence-313239-        9268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ribosome-tagged GCaMP (soma localized) expression in C. elegans ASH 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9268)
  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 9268)
  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: pAS19
FEATURES             Location/Qualifiers
     source          1..9268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(3923..3931,3932..3949,3950..3952,3953..3985,3986..3988,
                     3989..4012,4013..4039,4040..4096,4097..4102,4103..4372,
                     4373..4390,4391..4825,4826..4831,4832..5272)
                     /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      4019..4028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4309..4330
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4436..4457)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4697..4716)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(6785..6807)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6907..6926
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7008..7026)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7094..7198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7199..7267,7268..8059)
                     /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(7417..7436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8230..8818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8719..8738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8972..8989
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9106..9127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9142..9147,9148..9165,9166..9172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9180..9196
                     /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     9185..9207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9204..9220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9204..9220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttcagtgc ctacgtgcca gcattttttg tcgtagaggc atgtgcaggt tgccttaaaa
       61 gcacgcaaac attcttcgcc tacctggcag ttctcatcca gattattctg tcaaagaagt
      121 tgctttattt ttacgtgaac ctaaatacac attaattgat acagtgttcc accaaaattt
      181 ataaatttat tcttctttct ctcaagaaat cgtcatgcgt tatctttcca atttgctgtg
      241 cctaactctg acgtttatcg tttgctttgg cgccgagcgc gatgcagatg aagaagctta
      301 tgtactgatg aatcttctga aacaatctat tgatatggac caaactgaag ctcttccagg
      361 atttgtatcg agggatttca catatgatac ttgcaaggga atttggaata gaggtataca
      421 actgattatt agattttttc aaaatgaaat caattacaga taaatatatg gagtatctga
      481 aagtcttttc atctcgacca gacggtggta tgcaaccaat gtccactgat gtacctttct
      541 atctttctaa actttggcat ttgtatttta atttcagtgg aaggttacaa atgcacattt
      601 caacgacaat actctttatt tcaatattca caactacaat caacacttag caatctatcc
      661 tgatttggaa tttgaagcta aaaaagtgag catgaagaag gtagaggttt tctaaacttg
      721 gtttttcaga tcggtaaaga cgaacgttac attttgacaa gaggaaaaat tagcaaatgt
      781 gggacgttgg aaaaagaata ttaactttta gatataataa atcgaaattg aaatgtatct
      841 ttttatgcaa tgtacaatag aaatcatatg aagtctaaaa catgttagat agtatgctgc
      901 actataagga ttgaatctaa gtgtaagaaa tactcattat ttttagactt ttgagaacaa
      961 taatagtggt actgtttgaa ctgtgaccaa acttcagcaa attgaagttg tttaaagcat
     1021 gaagctgttc aaatccacga agctttgctc tccggtctta tttccaagta tattttataa
     1081 tttggcagat catctttttc tcagccaagt gttgaaatca tcgtgttcag aaggcatgtg
     1141 caatcccaat ctgattcgag cattgacgac cttaaatttg gcctttggta ttttgaaacg
     1201 catttaattg attcaagctt actttagagt tgttatctta cctcaatgta gttagtgttg
     1261 tagaacatat ttctatgagt gccccataaa ccagaaaagt caaacgaaaa tgtttcagat
     1321 atcaaaaaaa gtatcacaaa gcatgcgaaa aaaccaaatc tgaggtgcat ttgcgagggg
     1381 cacttcagat ttgaaacttt gaattttgca cctcgccatg tctataaaag gaggagtttg
     1441 ataatatctg acgcaagtag agtacaattt tattaatcca aataaattgt gccaattttt
     1501 ataagatgat ataaatgaga aacgggaatt gagaatatga tgagtagttt ttaaagtttg
     1561 tcattcaaaa ttgttgttgc ctgccatgca aatgaaagtc agaagagttc caaaccagaa
     1621 gaggaaatat gatttttcca tgtaacagta gttgtctttt tttagaacgg caggccaaac
     1681 acaaaagtat tccattattt cctgatttgc cttttctaaa caatttccag tcaagtgata
     1741 aattcacatt tccagtgctc tgaaaatctt gtcaaaaagt ttgtatttcg gatgtcaggt
     1801 gatgtttgta agacatacgt tagctggtct cttacgtatt ccaaaaccgc tacaatatga
     1861 taatcagtca aaatgaaaca ctaaagtttt tgaactgaat gatcacagag ttgattgaga
     1921 tagatagagt tataattttg aaggaatctt acgcatacgg aagcttcaaa gagtttagga
     1981 taatttcata gttagagtag acgtgacaaa gttattaata tcataatttg tttttctcga
     2041 aattgaatat gactgaaacg caagtcagag agattcctga acctaataat ctaaaatttt
     2101 aattgctatt ttcaatattg catttttcga ttcacgcgta aaggaagaga caatttcaca
     2161 gcatcggaac tatcttccga tttggaagat aattttttaa agtgttttcc ttattggctt
     2221 tgaaatttca ataaaagtac acctgacgaa ttttcaaata agtgaaaaat agaagcagac
     2281 atcgagtaaa cgtgcataaa ataaatgcaa taagtgctcg ttaagtaatg aaataaagtt
     2341 agtctacaaa ttgatcaacc agtacatgac taaaaattgg tgactgtttt gttttagtct
     2401 ttacccaatt tttatttcag cgatttcatt ttatactcaa atgtgatttt tctacatttt
     2461 ttcgaattta gaaaagatca gacgatttcc ttaaagagga tttctagttt agtctttgat
     2521 tcaataactt atagtacagc ttagttataa ctatagttca ctatgaaata tcacatttat
     2581 aatcgtttca tttaattgta aggacatttt tgcatcaaac gccagttttc tgtccccatt
     2641 taattataag ttttactcgg ccgatgcgag atggatgctc ctctcacaca ccttacccca
     2701 cgccttgaca ccttatttat tgggttttaa atttttttga tattttagag aattataaag
     2761 gtttccgtta aaaaaaacat ggccttcaag gcgagtacgt ctgcgttgta aactcacaaa
     2821 taaacttaaa aaatattttt ttttggattg aagctttatt tgaaaggata actcactcgc
     2881 tatctaaaaa ttgagaaagt tttgtaataa cacgaaattt catgaagtgc ggaaaatttt
     2941 tgaaaattgg atgaaactgt tcctttcacg agtttggtcg tgttttattc ttcattcaaa
     3001 taccaatgca acagtcggtt ttaacttttc aaaattatac ttcaagtttt tacaagtgca
     3061 tatgtaaaag tgttttctat cttcacacct tcgaaaacag caataataaa ttaaagccgc
     3121 gtgcagctat cttatagttt ggttctctgt attttcaaat atcttaagtg tcacatcaaa
     3181 ttcaatgata atttggaaat cacagaagat ctttaatgtg tttatagtct ctatgaactc
     3241 atcaattaca aatatatttc tagtcagact attcacacaa accgcagttt ttaatcatcc
     3301 atatcttctt ctcatggaag ttctagctaa cgtcaagtca gactttcaca agtatgagtt
     3361 ttattttgtc caacatttta tgctcttgat gaactcaaat atcatttctc tttaaactat
     3421 agtttcataa gattcagtaa ttttgattct gcagttcaga ttctaaattt atccgaagag
     3481 aaaaccaatt gatcaagtct ttattcaatt tccgagaaaa cgtggttgag cttaacacat
     3541 tttagaattt gtaaacgtgt tgttgttttt caaaacctcg ctaataattt ctccctagtt
     3601 actcccgtaa agtattttct ggaaaatttg caggtttaat agagttggtt tgaaatactg
     3661 caaatcaaaa cactttttat agattaaatc aattttgata atgttttctg tcacacataa
     3721 taaactgcac tcgcagaatt taatatttat tatttcagat tttgccattt ctaatttaag
     3781 ttgtgctgct ccggcgcgcc tctagaggat ccccgggatt ggccaaagga cccaaaggta
     3841 tgtttcgaat gatactaaca taacatagaa cattttcagg aggacccttg gctagcgtcg
     3901 acggtaccat ggtattgata tcatgggttc tcatcatcat catcatcatg gtatggctag
     3961 catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc
     4021 caccatggtc gactcatcac gtcgtaagtg gaataagaca ggtcacgcag tcagagctat
     4081 aggtcggctg agctcactcg agaacgtcta tatcaaggcc gacaagcaga agaacggcat
     4141 caaggcgaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca
     4201 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     4261 gagcgtgcag tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct
     4321 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac
     4381 cggagggagc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4441 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga
     4501 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4561 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4621 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg
     4681 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     4741 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     4801 cctggggcac aagctggagt acaacctgcc ggaccaactg actgaagagc agatcgcaga
     4861 atttaaagag gctttctccc tatttgacaa ggacggggat gggacaataa caaccaagga
     4921 gctggggacg gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat
     4981 gatcaatgaa gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat
     5041 gatggcaaga aaagggagct acagggacac ggaagaagaa attagagaag cgttcggtgt
     5101 gtttgataag gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa
     5161 ccttggagag aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga
     5221 tggggatggt caggtaaact acgaagagtt tgtacaaatg atgacagcga agtcaggccg
     5281 gactcagatc tcaagctcaa gcttcgagtt ctcgaaggtt tcccgcgagt ctcttaatga
     5341 ggctatcgct gaggtcctta agggatcctc ggaaaagcca cgcaagttcc gtgagactat
     5401 cgagctccag atcggtctca agaactacga cccacagaag gacaagcgtt tcagcggatc
     5461 catcagactg aagcacattc cacgcccaaa catgaaggtg tgcgtcttcg gagaccagca
     5521 ccatcttgac gaggctgccg ccggagacat tccatcgatg agcgccgatg acttgaagaa
     5581 gcttaacaag cagaagaagc tcatcaagaa gctcgccaag agctacgatg ctttcatcgc
     5641 ttccgaatcc ttgatcaagc agatcccacg tatcctcggt ccaggactca acaaggccgg
     5701 aaaattccct tccgtcgtta cccacggaga atctctccaa agcaagagtg acgagatccg
     5761 cgccaccgtc aagttccaga tgaagaaggt tctctgcctc tccgtcgccg tcggtcacgt
     5821 tggactcacc caggaggagc ttgtctccaa catctccctc tccatcaact tcctcgtctc
     5881 gctcttgaag aagaactggc agaacgtgag atcgttgaac atcaagtcca ccatgggaaa
     5941 gccacaaaga gtttattaag atatctgagc tccgcatcgg ccgctgtcat cagatcgcca
     6001 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     6061 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     6121 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     6181 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     6241 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     6301 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     6361 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     6421 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     6481 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     6541 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     6601 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     6661 tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa
     6721 ctgtttctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     6781 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6841 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     6901 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6961 acagatgcgt aaggagaaaa taccgcatca ggcggcctta attaagggcc tcgtgatacg
     7021 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     7081 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7141 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     7201 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     7261 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     7321 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     7381 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     7441 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     7501 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7561 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7621 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7681 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7741 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7801 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7861 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7921 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7981 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     8041 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     8101 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8161 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8221 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     8281 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     8341 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     8401 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     8461 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     8521 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8581 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8641 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8701 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8761 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8821 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     8881 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     8941 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     9001 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     9061 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     9121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     9181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
     9241 atgcgcggcc gcactgactg ggccggcc
//
