LOCUS       sequence-377515-        7680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Punc-25-GCaMP3::UrSL2 ::wCherry unc-54 3' UTR C.elegans DD neurou 
            expression of GCaMP3 RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7680)
  AUTHORS   Lu Y, Ahamed T, Mulcahy B, Meng J, Witvliet D, Guan SA, Holmyard D, 
            Hung W, Wen Q, Chisholm AD, Samuel ADT, Zhen M
  TITLE     Extrasynaptic signaling enables an asymmetric juvenile motor circuit
            to produce symmetric undulation.
  JOURNAL   Curr Biol. 2022 Nov 7;32(21):4631-4644.e5. doi: 
            10.1016/j.cub.2022.09.002. Epub 2022 Sep 30.
  PUBMED    36182701
REFERENCE   2  (bases 1 to 7680)
  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"; doi: 
            "10.1016/j.cub.2022.09.002"; journalName: "Curr Biol"; date: 
            "2022-11-7- 7"; volume: "32"; issue: "21"; pages: "4631-4644.e5"
COMMENT     Sequence Label: pJH2523
FEATURES             Location/Qualifiers
     source          1..7680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1409..1426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1430..1462
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1466..1474,1475..1489)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      1496..1505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1517..1573
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1836..1857
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2014..2035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2275..2294)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5120..5142)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5242..5261
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5339..5357)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5425..5529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5530..5598,5599..6390)
                     /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(5748..5767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6561..7149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7050..7069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7303..7320
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7437..7458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7473..7478,7479..7496,7497..7503)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7511..7527
                     /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     7516..7538
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7535..7551
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7535..7551
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcagcatgc ggccgcgaaa tatcgatttt ttccgagata tctaaaattg
       61 aaagaaattt tcgattttct gagaaaaata cgaaaaaatc gaaaaaaaaa cccaaatttc
      121 ggttttttcg taaaaaaagt cgtaaaaatg taattttttt cctgaaaaat cggaattttt
      181 tacaaaatat cggaaaatac tcaaaaaaaa gctgaaaatt tcgattttcc ggagaaaaaa
      241 tctttttaaa aaaatatttt ttttttcaga aaataagaaa agccgaaatt tataactatt
      301 tctccggaaa ttcgaaattt ttaacgaaat atcggaataa tttttagatt tttcaagttt
      361 gactttgcga gaaaaaaatc ggaaaaacct cgattcgacg ccgaaaaatg ctccttttcg
      421 aaaagatttt tgaaatttca gaaaatcgac atgcaagcgc gctccacggc gaaatgacaa
      481 cgatgatcca ccgccctcaa aaagttgggt ctcgttaggt atttggcggt aaaactggta
      541 aaactccagt tttgcctcca acgagaccca attttttggg gcggtggtgg agcgcgcttg
      601 cacaagctga aagcattttt ctgcgactcg ataatatttt gaaaacctgt gtcaattctc
      661 gaaattcttt ttttaaaaaa taatcccgag cttctctcag tcctcctcta tgaggatgtt
      721 cctttttttt ggtttttcaa ttttttttaa aattccaaat ttctgttgtg caattcactt
      781 ccccccaaga aatcccaaaa atccccagtt ttccccaaaa atgttccgtt ttcatgtgat
      841 ttttccccca tttttaaaac atttttttga ctttttttta aaatgattat tattattgtt
      901 ttctatttca tggccggtaa attatttttt ttctttcttt ttttttgctc ttttttttca
      961 agaattttcg aattgtttga agggctgctc atctaatctt ttgtcatttt gtcctgatgc
     1021 catcatttct gagaggacct ttgaagactc gtcacgaaac gggagggggg ctcaagtgag
     1081 cattattatt attattattg tcgcaaaaag tttaccccgg gctccctctg gctcccctct
     1141 ttgagcaagg gtttaagggc tcattttgat gacgaattgc tcattgggat tatagtcacg
     1201 cccctctttt ggagcaacta cacaactgag ccacagtaat ccttgggggc ggggtcagta
     1261 ggaccccctc cggaataggg aaaagctcag ttcaccgcca aaaattggcc aaaggaccca
     1321 aaggtatgtt tcgaatgata ctaacataac atagaacatt ttcaggagga cccttggcta
     1381 gcgtcgacgg atccaaaaaa tgggttctca tcatcatcat catcatggta tggctagcat
     1441 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atctcgccac
     1501 catggtcgac tcatcacgtc gtaagtggaa taagacaggt cacgcagtca gagctatagg
     1561 tcggctgagc tcactcgagg taagtttaaa cattatatac taactaacgg attatattta
     1621 aattttcaga acgtctatat caaggccgac aagcagaaga acggcatcaa ggcgaacttc
     1681 aagatccgcc acaacatcga ggacggcggc gtgcagctcg cctaccacta ccagcagaac
     1741 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cgtgcagtcc
     1801 aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1861 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg aggtaagttt
     1921 aaacagttcg gtactaacta accctgatta tttaaatttt cagggagcat ggtgagcaag
     1981 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2041 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     2101 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2161 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2221 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac
     2281 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2341 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2401 aacacgcgtg accaactgac tgaaggtaag tttaaacttg gacttactaa ctaaccatac
     2461 atatttaaat tttcagagca gatcgcagaa tttaaagagg ctttctccct atttgacaag
     2521 gacggggatg ggacaataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag
     2581 aaccccacag aagcagagct gcaggacatg atcaatgaag tagatgccga cggtgacggc
     2641 acaatcgact tccctgagtt cctgacaatg atggcaagaa aaatgaaaga cacagacagt
     2701 gaagaagaaa ttagagaagc gttccgtgtg tttgataagg atggcaatgg ctacatcagt
     2761 gcagcagagc ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt
     2821 gatgaaatga tcagggaagc agacatcgat ggggatggtc aggtaaacta cgaagagttt
     2881 gtacaaatga tgacagcgaa gtaggatccc cgagatcttc agcgctcagc attcgtagaa
     2941 ttgtaagttt aaactttctc atactaacta acgcgctcta tttaaatttt cagaattcgc
     3001 tgtctcatcc tactttcacc tagttaactg cttgtcttaa aatctatgct tctctttagt
     3061 atctaaaatt ttcctagaag cttacaagta tataaatggt ctcttctcaa taaaggttgt
     3121 atatttattc atcttattga atctgccatt tcctcgtttt tgcgagttta tataccttcc
     3181 aattttcttt ctattgtatt ttcaacttct aattttaatt cagggaaact gcttcaacgc
     3241 atcggtaccc tatggtctca aagggtgaag aagataacat ggcaattatt aaagagttta
     3301 tgcgtttcaa ggtgcatatg gagggatctg tcaatgggca tgagtttgaa attgaaggtg
     3361 aaggagaagg ccgaccatat gagggaacac aaaccgcaaa actaaaggta agtttaaaca
     3421 tatatatact aactaaccct gattatttaa attttcaggt aactaaaggc ggaccattac
     3481 cattcgcctg ggacatcctc tctccacagt tcatgtatgg aagtaaagct tatgttaaac
     3541 atccggcaga tataccagat tatttgaaac tttcattccc ggagggtttt aagtgggaac
     3601 gcgtaatgaa ttttgaagac ggaggagttg ttacagtgac gcaagactca aggtaagttt
     3661 aaacagttcg gtactaacta accatacata tttaaatttt cagcctccaa gatggagaat
     3721 ttatttataa agtcaaactt cgaggaacga atttcccctc ggatggatct gttatgcaga
     3781 agaagactat gggatgggaa gcttcaagtg aaagaatgta ccctgaagac ggtgctctta
     3841 agggagagat taaacaacgt cttaaattga aagatggagg acattacgat gctgaggtaa
     3901 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcaggtg aagacaactt
     3961 acaaagccaa aaaaccagtt cagctgccag gagcgtacaa tgttaatatt aaactggata
     4021 tcacctccca caacgaggat tacactatcg ttgagcaata tgaaagagct gaagggcggc
     4081 actcgacagg tggcatggat gaattgtata agtaggagct cccgagatct tcagcgctca
     4141 gcattcgtag aattgtaagt ttaaactttc tcatactaac taacgcgctc tatttaaatt
     4201 ttcagaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata
     4261 ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa cgagtaatat
     4321 ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca
     4381 tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact
     4441 tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta
     4501 gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc
     4561 attaataata attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt
     4621 tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     4681 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     4741 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     4801 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg
     4861 tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc
     4921 aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata
     4981 atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa
     5041 tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt
     5101 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     5161 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     5221 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5281 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     5341 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5401 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5461 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5521 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5581 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5641 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5701 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5761 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5821 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5881 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5941 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6001 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6061 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6121 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6181 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6241 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6301 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6361 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6421 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6481 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6541 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6601 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6661 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6721 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6781 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6841 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6901 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6961 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7021 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7081 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7141 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7201 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7261 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7321 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7381 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7441 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7501 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7561 gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga
     7621 gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg gaaatgaaat
//
