LOCUS       sequence-375288-        7395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptwk-40s-GCaMP::wScarlet unc-54 3' UTR C.elegans AVA/other neurons 
            expression GCaMP6s wScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7395)
  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 7395)
  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: pJH4260
FEATURES             Location/Qualifiers
     source          1..7395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    97..118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(133..138,139..156,157..163)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    171..187
                     /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     176..198
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     195..211
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     195..211
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1965..1982
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(2022..2030,2031..2045)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2073..2129
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(5175..5197)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5297..5316
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5394..5412)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5480..5584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5585..5653,5654..6445)
                     /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(5803..5822)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6616..7204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7105..7124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7358..7375
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
       61 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      121 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      181 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctgt aagtttaaac
      241 atgatcttac taactaacta ttctcattta aattttcaga gcttaaaaat ggctgaaatc
      301 actcacaacg atggatacgc taacaacttg gaaatgaaat aagcttgggc tgcagcatgc
      361 ggccgctcta atcactatca cgtgggatct ggataaattt tctccaaatt tctcataaaa
      421 aatggaaaaa aagaaaagga aatccggttt gtgtatttcc attttcaatt tgatgcatta
      481 aaccagaact tcgttaattt cccttctctt ctccctcaag cgtgcacact ttttttagat
      541 gagaagggct ctatttcgat tggttgtttt tgttttgaac cagatttttt tttcacaaaa
      601 ataaaatttg gaaccttttg ccagtttaat aaagtttttt gttgggaatg ttcctaaaaa
      661 tttttaattt atctaatata aaacttgata gaaacattaa ttaacctgct tttttgcatt
      721 tatttccaac tgaattaaaa taaatgtgaa ttttttttac tatgtaaata aatcgaaaat
      781 tctggttcct gattctcaag accgcatctc aacttttcag ccggtaattt gtccgagagg
      841 agtccaattt tcaaattttt gatcaaaatg tctactttga tcgacatcct ccttcttctg
      901 caaggtgtgt atcatcgtct tgaaatttca aattccctaa gggaacgaaa agcttgtctc
      961 ttttggcaga ttcgattcga tttcacctct ttctcccccc tcctgcctac aatgttgaaa
     1021 gggtcccctc cttgtgctcc aaattgctca taaactcctc catactggac gcttcttttt
     1081 gcggaaattg gagtcccaga tatattggaa gagataaacg gttttcgggt ttgcagttgt
     1141 tgccattttc actgattttt attcaagtag tactacttga agttttgttc ctgagtaaaa
     1201 ttttataggt tccatgattt atgaaattgt tctatagtag caccaaactt ttcagaactt
     1261 ttttgttttt tttttcaatt ttacagtaaa aagttattct caaatgtttt ctcaattttt
     1321 gccccttatt aactcacgta cgtatgtagt ttacagagcg tatgtagatg tagttccagt
     1381 ttttggcact tataatttta gaagtcgacc gataataaat gtttaggcat gccaaatcat
     1441 cataagctac tcaaaaaata gaaaattttt gatatgtagc aattttctaa aactttatct
     1501 aaaaatcttt ttttccaaaa aatgcctttg gttaataaat tgtaaatcaa aattcagcaa
     1561 caaaaaacct cactggaaaa gtgggagtaa aaatacgaaa taaaattttc gtaacttttt
     1621 gtcaaaacag aatcttaaat ttcccactat acccatttgt ccttcaaaaa tgccatttaa
     1681 agtaagcaaa gaggtatcat aaattctcac agaacgtatg tagtttttgt gtcctttact
     1741 tctccattgc ctcatatttt cacttttttc gtggccgcag ctcacagtgc acacatcaca
     1801 gaatgttcca gagccgaatg attttggatg gaatatcgag tgatgaatat tcacccggga
     1861 ttggccaaag gacccaaagg tatgtttcga atgatactaa cataacatag aacattttca
     1921 ggaggaccct tggctagcgt cgacggatcc aaaaaatggg atctcaccac caccaccacc
     1981 acggaatggc ctccatgacc ggaggacagc agatgggacg tgacctctac gacgacgacg
     2041 acaaggacct cgccactatg gtcgactcct cccgtcgtaa gtggaacaag accggacacg
     2101 ccgtccgtgc catcggacgt ctctcctccc tcgagaacgt ctacatcaag gccgacaagc
     2161 agaagaacgg aatcaaggta agtttaaaca tgattttact aactaactaa tctgatttaa
     2221 attttcaggc caacttccac atccgtcaca acatcgagga cggaggagtc cagctcgcct
     2281 accactacca gcagaacacc ccaatcggag acggaccagt cctcctccca gacaaccact
     2341 acctctccgt ccagtccaag ctctccaagg acccaaacga gaagcgtgac cacatggtcc
     2401 tccttgagtt cgtcaccgcc gccggaatca ccctcggaat ggacgagctc tacaagggag
     2461 gaaccggagg ttccatggtc tccaagggag aggagctctt caccggagtc gtcccaatcc
     2521 tcgtcgagct cgacggagac gtcaacggac acaagttctc cgtctctgga gaggtaagtt
     2581 taaacatgat tttactaact aactaatctg atttaaattt tcagggagag ggagacgcca
     2641 cctacggaaa gctcaccctc aagttcatct gcaccaccgg aaagctccca gtcccgtggc
     2701 cgaccctcgt caccaccctc acctacggag tccagtgctt ctcccgttac ccagaccaca
     2761 tgaagcagca cgacttcttc aagtccgcca tgccagaggg atacatccag gagcgtacca
     2821 tcttcttcaa ggacgacgga aactacaaga cccgtgccga ggtcaagttc gagggagaca
     2881 ccctcgtcaa ccgtatcgag ctcaagggaa tcgacttcaa ggaggacgga aacatcctcg
     2941 gacacaagct tgagtacaac ctcccagacc agctcaccga ggagcagatc gccgagttca
     3001 aggtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt caggaggcct
     3061 tctccctctt cgacaaggac ggagacggaa ccatcaccac caaggagctc ggaaccgtca
     3121 tgcgttccct cggacagaac ccaaccgagg ccgagctcca ggacatgatc aacgaggtcg
     3181 acgccgacgg agacggaacc atcgacttcc cagagttcct caccatgatg gcccgtaaga
     3241 tgaagtaccg tgacaccgag gaggagatcc gtgaggcctt cggagtcttc gacaaggacg
     3301 gaaacggata catctccgcc gccgagctcc gtcacgtcat gaccaacctc ggagagaagc
     3361 tcaccgacga ggaggtcgac gagatgatcc gtgaggccga catcgacgga gacggacagg
     3421 tcaactacga ggagttcgtc cagatgatga ccgccaaggg aggacgtgga ggatctggcg
     3481 gcgcgccaaa aatggtcagc aagggagagg cagttatcaa ggagttcatg cgtttcaagg
     3541 tccacatgga gggatccatg aacggacacg agttcgagat cgagggagag ggagagggac
     3601 gtccatacga gggaacccaa accgccaagc tcaaggtcac caagggagga ccactcccat
     3661 tctcctggga catcctctcc ccacaattca tgtacggatc ccgtgccttc accaagcacc
     3721 cagccgacat cccagactac tacaagcaat ccttcccaga gggattcaag tgggagcgtg
     3781 tcatgaactt cgaggacgga ggagccgtca ccgtcaccca agacacctcc ctcgaggacg
     3841 gaaccctcat ctacaaggtc aagctccgtg gaaccaactt cccaccagac ggaccagtca
     3901 tgcaaaagaa gaccatggga tgggaggcct ccaccgagcg tctctaccca gaggacggag
     3961 tcctcaaggg agacatcaag atggccctcc gtctcaagga cggaggacgt tacctcgccg
     4021 acttcaagac cacctacaag gccaagaagc cagtccaaat gccaggagcc tacaacgtcg
     4081 accgtaagct cgacatcacc tcccacaacg aggactacac cgtcgtcgag caatacgagc
     4141 gttccgaggg acgtcactcc accggaggaa tggacgagct ctacaagtaa gctcagcatt
     4201 cgtagaattg taagtttaaa ctttctcata ctaactaacg cgctctattt aaattttcag
     4261 aattccaact gagcgccggt cgctaccatt accaacttgt ctggtgtcaa aaataatagg
     4321 ggccgctgtc atcagagtaa gtttaaactg agttctacta actaacgagt aatatttaaa
     4381 ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct
     4441 acatgctctt tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaacttctct
     4501 taatttcttt gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc
     4561 aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa
     4621 taataattct atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttactt
     4681 ctgataaatt tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt
     4741 tacgtttcag tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt
     4801 caaatcatgc tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa
     4861 agtttatgaa attaattttc ctgcttttgc tttttggggt ttcccctatt gtttgtcaag
     4921 atttcgagga cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa
     4981 agatcggaag aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg
     5041 aaagtggagt agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac
     5101 caaacataac tgtttcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt
     5161 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5221 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5281 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5341 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt
     5401 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5461 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5521 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5581 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5641 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5701 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5761 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5821 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5881 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5941 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6001 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6061 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6121 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6181 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6241 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6301 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6361 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6421 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6481 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6541 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6601 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6661 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6721 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6781 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6841 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6901 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6961 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7021 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7081 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7141 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7201 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7261 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7321 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7381 ggaagagcgc ccaat
//
