LOCUS       sequence-408444-        7539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptwk-40s GCaMP6s::mNeptune unc-54 3' UTR C.elegans AVA,AVB and other
            neurons expression of GCaMP6 mNeptune.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7539)
  AUTHORS   Meng J, Ahamed T, Yu B, Hung W, Ei Mouridi S, Wang Z, Zhang Y, Wen 
            Q, Boulin T, Gao S, Zhen M
  TITLE     A tonically active master neuron modulates mutually exclusive motor 
            states at two timescales.
  JOURNAL   Sci Adv. 2024 Apr 12;10(15):eadk0002. doi: 10.1126/sciadv.adk0002. 
            Epub 2024 Apr 10.
  PUBMED    38598630
REFERENCE   2  (bases 1 to 7539)
  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: "Sci Adv."; 
            date: "2024-04-12"; volume: "10(15):eadk0002. doi"; pages: " 
            10.1126/sciadv.adk0002. Epub 2024 Apr 10"
COMMENT     Sequence Label: pJH4293
FEATURES             Location/Qualifiers
     source          1..7539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1625..1642
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(1682..1690,1691..1705)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1733..1789
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(4979..5001)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5101..5120
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5198..5216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5284..5388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5389..5457,5458..6249)
                     /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(5607..5626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6420..7008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6909..6928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7162..7179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7296..7317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7332..7337,7338..7355,7356..7362)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7370..7386
                     /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     7375..7397
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7394..7410
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7394..7410
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcagcatgc ggccgctcta atcactatca cgtgggatct ggataaattt
       61 tctccaaatt tctcataaaa aatggaaaaa aagaaaagga aatccggttt gtgtatttcc
      121 attttcaatt tgatgcatta aaccagaact tcgttaattt cccttctctt ctccctcaag
      181 cgtgcacact ttttttagat gagaagggct ctatttcgat tggttgtttt tgttttgaac
      241 cagatttttt tttcacaaaa ataaaatttg gaaccttttg ccagtttaat aaagtttttt
      301 gttgggaatg ttcctaaaaa tttttaattt atctaatata aaacttgata gaaacattaa
      361 ttaacctgct tttttgcatt tatttccaac tgaattaaaa taaatgtgaa ttttttttac
      421 tatgtaaata aatcgaaaat tctggttcct gattctcaag accgcatctc aacttttcag
      481 ccggtaattt gtccgagagg agtccaattt tcaaattttt gatcaaaatg tctactttga
      541 tcgacatcct ccttcttctg caaggtgtgt atcatcgtct tgaaatttca aattccctaa
      601 gggaacgaaa agcttgtctc ttttggcaga ttcgattcga tttcacctct ttctcccccc
      661 tcctgcctac aatgttgaaa gggtcccctc cttgtgctcc aaattgctca taaactcctc
      721 catactggac gcttcttttt gcggaaattg gagtcccaga tatattggaa gagataaacg
      781 gttttcgggt ttgcagttgt tgccattttc actgattttt attcaagtag tactacttga
      841 agttttgttc ctgagtaaaa ttttataggt tccatgattt atgaaattgt tctatagtag
      901 caccaaactt ttcagaactt ttttgttttt tttttcaatt ttacagtaaa aagttattct
      961 caaatgtttt ctcaattttt gccccttatt aactcacgta cgtatgtagt ttacagagcg
     1021 tatgtagatg tagttccagt ttttggcact tataatttta gaagtcgacc gataataaat
     1081 gtttaggcat gccaaatcat cataagctac tcaaaaaata gaaaattttt gatatgtagc
     1141 aattttctaa aactttatct aaaaatcttt ttttccaaaa aatgcctttg gttaataaat
     1201 tgtaaatcaa aattcagcaa caaaaaacct cactggaaaa gtgggagtaa aaatacgaaa
     1261 taaaattttc gtaacttttt gtcaaaacag aatcttaaat ttcccactat acccatttgt
     1321 ccttcaaaaa tgccatttaa agtaagcaaa gaggtatcat aaattctcac agaacgtatg
     1381 tagtttttgt gtcctttact tctccattgc ctcatatttt cacttttttc gtggccgcag
     1441 ctcacagtgc acacatcaca gaatgttcca gagccgaatg attttggatg gaatatcgag
     1501 tgatgaatat tcacccggga ttggccaaag gacccaaagg tatgtttcga atgatactaa
     1561 cataacatag aacattttca ggaggaccct tggctagcgt cgacggatcc aaaaaatggg
     1621 atctcaccac caccaccacc acggaatggc ctccatgacc ggaggacagc agatgggacg
     1681 tgacctctac gacgacgacg acaaggacct cgccactatg gtcgactcct cccgtcgtaa
     1741 gtggaacaag accggacacg ccgtccgtgc catcggacgt ctctcctccc tcgagaacgt
     1801 ctacatcaag gccgacaagc agaagaacgg aatcaaggta agtttaaaca tgattttact
     1861 aactaactaa tctgatttaa attttcaggc caacttccac atccgtcaca acatcgagga
     1921 cggaggagtc cagctcgcct accactacca gcagaacacc ccaatcggag acggaccagt
     1981 cctcctccca gacaaccact acctctccgt ccagtccaag ctctccaagg acccaaacga
     2041 gaagcgtgac cacatggtcc tccttgagtt cgtcaccgcc gccggaatca ccctcggaat
     2101 ggacgagctc tacaagggag gaaccggagg ttccatggtc tccaagggag aggagctctt
     2161 caccggagtc gtcccaatcc tcgtcgagct cgacggagac gtcaacggac acaagttctc
     2221 cgtctctgga gaggtaagtt taaacatgat tttactaact aactaatctg atttaaattt
     2281 tcagggagag ggagacgcca cctacggaaa gctcaccctc aagttcatct gcaccaccgg
     2341 aaagctccca gtcccgtggc cgaccctcgt caccaccctc acctacggag tccagtgctt
     2401 ctcccgttac ccagaccaca tgaagcagca cgacttcttc aagtccgcca tgccagaggg
     2461 atacatccag gagcgtacca tcttcttcaa ggacgacgga aactacaaga cccgtgccga
     2521 ggtcaagttc gagggagaca ccctcgtcaa ccgtatcgag ctcaagggaa tcgacttcaa
     2581 ggaggacgga aacatcctcg gacacaagct tgagtacaac ctcccagacc agctcaccga
     2641 ggagcagatc gccgagttca aggtaagttt aaacatgatt ttactaacta actaatctga
     2701 tttaaatttt caggaggcct tctccctctt cgacaaggac ggagacggaa ccatcaccac
     2761 caaggagctc ggaaccgtca tgcgttccct cggacagaac ccaaccgagg ccgagctcca
     2821 ggacatgatc aacgaggtcg acgccgacgg agacggaacc atcgacttcc cagagttcct
     2881 caccatgatg gcccgtaaga tgaagtaccg tgacaccgag gaggagatcc gtgaggcctt
     2941 cggagtcttc gacaaggacg gaaacggata catctccgcc gccgagctcc gtcacgtcat
     3001 gaccaacctc ggagagaagc tcaccgacga ggaggtcgac gagatgatcc gtgaggccga
     3061 catcgacgga gacggacagg tcaactacga ggagttcgtc cagatgatga ccgccaaggg
     3121 aggacgtgga ggatctggcg gcgcgccaaa aatgcgcgct gtctccaagg gagaggagct
     3181 catcaaggag aacatgcaca ccaagctcta catggaggga accgtcaaca accaccactt
     3241 caagtgcacc cacgaagggg agggcaagcc ctacgagggc acccagacca acaggattaa
     3301 ggtggtggag ggaggccccc tgccgttcgc attcgacatc ctggccacct gctttatgta
     3361 cgggagcaag accttcatca accacaccca aggaatccca gacttcttca agcaatcctt
     3421 cgtaagttta aacatgattt tactaactaa ctaatctgat ttaaattttc agcctgaggg
     3481 cttcacatgg gagagagtca ccacatacga agacgggggc gtgcttaccg ttacccagga
     3541 caccagcctc caggacggct gcttgatcta caacgtcaag ctcagagggg tgaacttccc
     3601 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccagta ccgagaccct
     3661 gtaccccgct gacggcggcc tggaaggcag atgcgacatg gccctgaagc tcgtgggcgg
     3721 gggccacctg cactgcaacc tgaagaccac atacagatcc aagaaacccg ctaagaacct
     3781 caagatgcca ggagtaagtt taaacatata tatactaact aaccctgatt atttaaattt
     3841 tcaggtctac tttgtggacc gcagactgga aagaatcaag gaggccgaca atgagaccta
     3901 cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca
     3961 caaacttaat ggcatggacg agctgtacaa gtaagctcag cattcgtaga attgtaagtt
     4021 taaactttct catactaact aacgcgctct atttaaattt tcagaattcc aactgagcgc
     4081 cggtcgctac cattaccaac ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga
     4141 gtaagtttaa actgagttct actaactaac gagtaatatt taaattttca gcatctcgcg
     4201 cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc
     4261 tgtgctccca ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt
     4321 tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt
     4381 cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca
     4441 aaatctacac aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg
     4501 aaacatcata gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg
     4561 accgcaattt ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg
     4621 tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat
     4681 tttcctgctt ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt
     4741 ttttcttgct aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt
     4801 tgggtttgag gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc
     4861 tatggggttt ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc
     4921 ctactagtcg gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4981 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5041 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5101 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     5161 gcgtaaggag aaaataccgc atcaggcggc cttaagggcc tcgtgatacg cctattttta
     5221 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5281 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5341 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5401 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5461 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5521 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5581 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5641 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5701 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5761 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5821 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5881 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5941 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6001 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6061 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6121 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6181 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6241 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6301 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6361 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6421 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6481 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6541 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6601 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6661 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6721 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6781 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6841 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6901 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6961 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7021 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7081 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7141 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7201 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7261 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7321 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7381 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctgta agtttaaaca
     7441 tgatcttact aactaactat tctcatttaa attttcagag cttaaaaatg gctgaaatca
     7501 ctcacaacga tggatacgct aacaacttgg aaatgaaat
//
