LOCUS       sequence-377552-        7363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Punc-25-GCaMP3 ::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 7363)
  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 7363)
  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: pJH2999
FEATURES             Location/Qualifiers
     source          1..7363
                     /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(4803..4825)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4925..4944
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5022..5040)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5108..5212
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5213..5281,5282..6073)
                     /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(5431..5450)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6244..6832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6733..6752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6986..7003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7120..7141
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7156..7161,7162..7179,7180..7186)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7194..7210
                     /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     7199..7221
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7218..7234
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7218..7234
                     /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 gggaggacgt ggaggatctg gcggcgcgcc aaaaatggtc
     2941 tcaaagggtg aagaagataa catggcaatt attaaagagt ttatgcgttt caaggtgcat
     3001 atggagggat ctgtcaatgg gcatgagttt gaaattgaag gtgaaggaga aggccgacca
     3061 tatgagggaa cacaaaccgc aaaactaaag gtaagtttaa acatatatat actaactaac
     3121 cctgattatt taaattttca ggtaactaaa ggcggaccat taccattcgc ctgggacatc
     3181 ctctctccac agttcatgta tggaagtaaa gcttatgtta aacatccggc agatatacca
     3241 gattatttga aactttcatt cccggagggt tttaagtggg aacgcgtaat gaattttgaa
     3301 gacggaggag ttgttacagt gacgcaagac tcaaggtaag tttaaacagt tcggtactaa
     3361 ctaaccatac atatttaaat tttcagcctc caagatggag aatttattta taaagtcaaa
     3421 cttcgaggaa cgaatttccc ctcggatgga tctgttatgc agaagaagac tatgggatgg
     3481 gaagcttcaa gtgaaagaat gtaccctgaa gacggtgctc ttaagggaga gattaaacaa
     3541 cgtcttaaat tgaaagatgg aggacattac gatgctgagg taagtttaaa catgatttta
     3601 ctaactaact aatctgattt aaattttcag gtgaagacaa cttacaaagc caaaaaacca
     3661 gttcagctgc caggagcgta caatgttaat attaaactgg atatcacctc ccacaacgag
     3721 gattacacta tcgttgagca atatgaaaga gctgaagggc ggcactcgac aggtggcatg
     3781 gatgaattgt ataagtagga gctcccgaga tcttcagcgc tcagcattcg tagaattgta
     3841 agtttaaact ttctcatact aactaacgcg ctctatttaa attttcagaa ttccaactga
     3901 gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat
     3961 cagagtaagt ttaaactgag ttctactaac taacgagtaa tatttaaatt ttcagcatct
     4021 cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc
     4081 tccctgtgct cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt
     4141 tttttagctt cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt
     4201 aattcgtaat aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat
     4261 cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt
     4321 tttgaaacat catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt
     4381 tatgaccgca atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc
     4441 atcgtgaaaa agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat
     4501 taattttcct gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg
     4561 gcgtttttct tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa
     4621 ggtttgggtt tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag
     4681 tgtctatggg gtttttgcct taaatgacag aatacattcc caatatacca aacataactg
     4741 tttcctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     4801 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4861 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     4921 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4981 agatgcgtaa ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt
     5041 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5101 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5161 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5221 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5281 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5341 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5401 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5461 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5521 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5581 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5641 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5701 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5761 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5821 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5881 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5941 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6001 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6061 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6121 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6181 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6241 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6301 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6361 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6421 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6481 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6541 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6601 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6661 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6721 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6781 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6841 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6901 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6961 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7021 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7081 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7141 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7201 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tgtaagttta
     7261 aacatgatct tactaactaa ctattctcat ttaaattttc agagcttaaa aatggctgaa
     7321 atcactcaca acgatggata cgctaacaac ttggaaatga aat
//
