LOCUS       sequence-313236-        8528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ribosome-tagged GCaMP (soma localized) expression in C. elegans AFD 
            neurons (and others).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8528)
  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 8528)
  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: pAS16
FEATURES             Location/Qualifiers
     source          1..8528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(3183..3191,3192..3209,3210..3212,3213..3245,3246..3248,
                     3249..3272,3273..3299,3300..3356,3357..3362,3363..3632,
                     3633..3650,3651..4085,4086..4091,4092..4532)
                     /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      3279..3288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3569..3590
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3696..3717)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3957..3976)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(6045..6067)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6167..6186
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6268..6286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6354..6458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6459..6527,6528..7319)
                     /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(6677..6696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7490..8078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7979..7998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8232..8249
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8366..8387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8402..8407,8408..8425,8426..8432)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8440..8456
                     /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     8445..8467
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8464..8480
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8464..8480
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tcatgtttgc ataaactcca aaattaaaga catacccgag ggactttttt tttctataga
       61 aaaccataaa actggaagtg tgaaagatat gtctggtccg acatctaccg ggccccgcgc
      121 cgcactgtgt taaccctgca aaaccagtct agggaaccag gtaaatgtcg gaccatacaa
      181 tttagtcact cttcctattt tgagactttc tcaaaaattg tttctcagaa tgaacgtttc
      241 cgaaatttag aagtttctgc atgcactttt gttgtgttga gataaagttt tcgaaatttt
      301 cgtgttttgt tttgctttac attttcttta tttgaaagtg ctggaaattc ttgcagaaat
      361 gaaagaaaga aaaattcaaa tttacaaaaa tatatttaaa gcttgcatgc agtactaatt
      421 gaacgaagga atggtaagac gccaattatt gagttattta acagaagtaa gtaacgatta
      481 gtgtactttt ctaatgaaac aaaaactata agatagatag aaataaaaga agcaacacca
      541 caaccacact ttcagttaga tgccatgaga tcgttttgaa agtagataat tgcgtttaga
      601 aatatgatcc tgttggctaa agccaataag catggagggg tgcgaggagg ggaccagaaa
      661 aatattcctt ctatcttttt ttgcttgcag aaaattttga agaacaaggg ccaaatcttt
      721 aaagcagggg gaagagtgga gaggatggga gggcaacgag acaaaaattc atgcacaacg
      781 agcgagaaaa gtcccttacg cgtgaaaaaa atgtggagca acaaggtctt accgaaaaaa
      841 ggcactccga aaaaaacggc atttcagctg acgggtgaag atagaatacg caacaatcta
      901 tcaagttgtt cagaagaaga aaaaaacgct gatgcccatt ggaaatgtgt tacgcgttta
      961 cgcgtctaac agtcatcaaa acacgataaa ctcgagattg agtgctcaaa acggatttgt
     1021 tggaaaaatt gggagaaaag tggggaaaag taacggtgac ccagatgcgg cagtttgaaa
     1081 gatgtagcct aatggaatta gagcaatctt ctagtagtac ggaattggtc gctttaaagt
     1141 acgcagttag tctctgcaga actaaaagac aaaaagaggc acgaatctcg gagaagacac
     1201 aagaagggcc cctcctctta agacaacaca atttttttgg aacaacggaa cttgttgctg
     1261 gcaataatga ttgtagaaaa tctctaattt atgagcatgt aacacattga cgtgctcgta
     1321 aaagcaacaa gtataccacc aaagcagcag ataatgccag ttgactccag atgacaaatc
     1381 ggtgtcgtga agtcgcggca atcccccaga gcaaatgccg catcttttca aacacttacc
     1441 aatgtgcgct ccaaatttga tgaattagct atttggcgta tattcgaaac tggagaatgt
     1501 tcactttgcc gttttttgtt gttttaattt tatgtcagaa gtctgtagat tctgagtgag
     1561 tgaatgagtt gtaaaacttt tacatcaatg aacttctatg taatctggat gcggatttag
     1621 tatattttga gtaacatttt tgtttttttg tacttccttt tttctcaatt ttcgtaatct
     1681 cttttcatat gattttttat aacacgtcac aattgtttca ttgcaaacta cagattcata
     1741 ttgtgagttt ctattcaata acgacaaaaa cattttccaa acagatgccc taaattttga
     1801 cactttttgc ttcacttctg ctcataaaaa acaccttgta acctgatttt gaattttgtt
     1861 gggaacttgt gaattattga aaccttcgga agtttttggc aatcgacgaa gtctgcaatt
     1921 cttctgaaag tgtcaaaaaa taattttttt ttgatttgaa gtgtatttca aaaatctaaa
     1981 aaaaaacggt gacagcaaaa tacacgtttt acgttttatt ttcgacaatt cttatatcct
     2041 tatatccata tatccattgg cagatttggg aactcctggt tgtaaaatat ctggaaaaaa
     2101 gaaaattctg ccaaaattgc taaaattgtg tgagggtata ttacttctta gaaatcgtac
     2161 cccgaatgtt tctataacat cctaataatt atttctttgt catgtgccaa ttggttttat
     2221 tatttagacg aattttgtat gtgattttga gatagctgca ttaaacaaaa gatgagtata
     2281 aaccgatcag ctcagccagg ttaacagtac ggctctgtcc gtaaccatct ggaaaaaact
     2341 tgtaaagttc atcgagttac ccggaattgt caaagctttg caaaagcttg gagggttgaa
     2401 attgcaattc cctcgttttt ttatggcact tcaatatttt gaaaaaaaac caaattttaa
     2461 cagaaacctt caattttgtc ccgtgtcata gttggctttg cttaaattat tggtaaacga
     2521 ccgaaacatg caatttaaaa atgacagttt aaaacagtaa atttcagaca gctgcgacac
     2581 ggaaaacaaa ccgccgtaca acaaagttga acaattacta ttttcgatta tttgagcaca
     2641 ttttgagttt acacgttcaa aattcccaga accgttactg atcctttgaa ataatttagt
     2701 gcattttttt tgaaattatt tttcaaatca aatcttagtc acgtcacttg aattgataac
     2761 ttctgtcaaa cccacgtcct gacttgtttt gacacacatc ccaagaaact gtcactaaat
     2821 ccgtgtttca gagctttcaa tatgaaagaa aaaagctagt catatacatg aaggacgcgt
     2881 gagtcgactt atcgccaaat aacaagacgc gcggatttca gtcgtaataa agcggtttaa
     2941 gtcgacgatt atcagcgggg cggagctgcg atgaaatcca aaatccaggc gaggaggaaa
     3001 gagccattcg ttgtcattca atcttcactt tccattccaa ctagggctcc tcggcgcgcc
     3061 tctagaggat ccccgggatt ggccaaagga cccaaaggta tgtttcgaat gatactaaca
     3121 taacatagaa cattttcagg aggacccttg gctagcgtcg acggtaccat ggtattgata
     3181 tcatgggttc tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa
     3241 tgggtcggga tctgtacgac gatgacgata aggatctcgc caccatggtc gactcatcac
     3301 gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg agctcactcg
     3361 agaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac ttcaagatcc
     3421 gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca
     3481 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag tccaaacttt
     3541 cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3601 ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc atggtgagca
     3661 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3721 acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga
     3781 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     3841 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     3901 tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg
     3961 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4021 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4081 acaacctgcc ggaccaactg actgaagagc agatcgcaga atttaaagag gctttctccc
     4141 tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg gtgatgcggt
     4201 ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg
     4261 acggtgacgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga aaagggagct
     4321 acagggacac ggaagaagaa attagagaag cgttcggtgt gtttgataag gatggcaatg
     4381 gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag aagttaacag
     4441 atgaagaggt tgatgaaatg atcagggaag cagacatcga tggggatggt caggtaaact
     4501 acgaagagtt tgtacaaatg atgacagcga agtcaggccg gactcagatc tcaagctcaa
     4561 gcttcgagtt ctcgaaggtt tcccgcgagt ctcttaatga ggctatcgct gaggtcctta
     4621 agggatcctc ggaaaagcca cgcaagttcc gtgagactat cgagctccag atcggtctca
     4681 agaactacga cccacagaag gacaagcgtt tcagcggatc catcagactg aagcacattc
     4741 cacgcccaaa catgaaggtg tgcgtcttcg gagaccagca ccatcttgac gaggctgccg
     4801 ccggagacat tccatcgatg agcgccgatg acttgaagaa gcttaacaag cagaagaagc
     4861 tcatcaagaa gctcgccaag agctacgatg ctttcatcgc ttccgaatcc ttgatcaagc
     4921 agatcccacg tatcctcggt ccaggactca acaaggccgg aaaattccct tccgtcgtta
     4981 cccacggaga atctctccaa agcaagagtg acgagatccg cgccaccgtc aagttccaga
     5041 tgaagaaggt tctctgcctc tccgtcgccg tcggtcacgt tggactcacc caggaggagc
     5101 ttgtctccaa catctccctc tccatcaact tcctcgtctc gctcttgaag aagaactggc
     5161 agaacgtgag atcgttgaac atcaagtcca ccatgggaaa gccacaaaga gtttattaag
     5221 atatctgagc tccgcatcgg ccgctgtcat cagatcgcca tctcgcgccc gtgcctctga
     5281 cttctaagtc caattactct tcaacatccc tacatgctct ttctccctgt gctcccaccc
     5341 cctatttttg ttattatcaa aaaacttctc ttaatttctt tgttttttag cttcttttaa
     5401 gtcacctcta acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt
     5461 cgaaaaaaat tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat
     5521 gttctgtgta cacttcttat gttttttact tctgataaat ttttttgaaa catcatagaa
     5581 aaaaccgcac acaaaatacc ttatcatatg ttacgtttca gtttatgacc gcaattttta
     5641 tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg
     5701 gagtattttt ggaatttttc aatcaagtga aagtttatga aattaatttt cctgcttttg
     5761 ctttttgggg tttcccctat tgtttgtcaa gatttcgagg acggcgtttt tcttgctaaa
     5821 atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct
     5881 cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat ggggtttttg
     5941 ccttaaatga cagaatacat tcccaatata ccaaacataa ctgtttctac tagtcggccg
     6001 tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6061 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6121 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6181 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6241 taccgcatca ggcggcctta attaagggcc tcgtgatacg cctattttta taggttaatg
     6301 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     6361 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6421 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6481 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6541 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6601 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6661 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6721 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6781 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6841 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6901 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6961 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     7021 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     7081 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7141 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     7201 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7261 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7321 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7381 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7441 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7501 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7561 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7621 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7681 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7741 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7801 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7861 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7921 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7981 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8041 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     8101 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     8161 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     8221 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     8281 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     8341 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     8401 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     8461 acacaggaaa cagctatgac catgattacg ccaagcttgc atgcgcggcc gcactgactg
     8521 ggccggcc
//
