LOCUS       sequence-412786-        7734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptwk-40s GCaMP6s T2A NLS::mNeptune unc-54 3' UTR C.elegans AVA,AVB 
            and other neurons expression of GCaMP6 and nuclear mNeptune.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7734)
  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 7734)
  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: pJH4542
FEATURES             Location/Qualifiers
     source          1..7734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1328..1384
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1400..1420
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1445..1465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1487..1507
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3347..3369)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3469..3488
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3566..3584)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3652..3756
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3757..3825,3826..4617)
                     /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(3975..3994)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4788..5376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5277..5296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5530..5547
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5664..5685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5700..5705,5706..5723,5724..5730)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5738..5754
                     /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     5743..5765
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5762..5778
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5762..5778
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             7532..7549
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(7589..7597,7598..7612)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             7640..7696
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
ORIGIN
        1 cggaatcaag gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca
       61 ggccaacttc cacatccgtc acaacatcga ggacggagga gtccagctcg cctaccacta
      121 ccagcagaac accccaatcg gagacggacc agtcctcctc ccagacaacc actacctctc
      181 cgtccagtcc aagctctcca aggacccaaa cgagaagcgt gaccacatgg tcctccttga
      241 gttcgtcacc gccgccggaa tcaccctcgg aatggacgag ctctacaagg gaggaaccgg
      301 aggttccatg gtctccaagg gagaggagct cttcaccgga gtcgtcccaa tcctcgtcga
      361 gctcgacgga gacgtcaacg gacacaagtt ctccgtctct ggagaggtaa gtttaaacat
      421 gattttacta actaactaat ctgatttaaa ttttcaggga gagggagacg ccacctacgg
      481 aaagctcacc ctcaagttca tctgcaccac cggaaagctc ccagtcccgt ggccgaccct
      541 cgtcaccacc ctcacctacg gagtccagtg cttctcccgt tacccagacc acatgaagca
      601 gcacgacttc ttcaagtccg ccatgccaga gggatacatc caggagcgta ccatcttctt
      661 caaggacgac ggaaactaca agacccgtgc cgaggtcaag ttcgagggag acaccctcgt
      721 caaccgtatc gagctcaagg gaatcgactt caaggaggac ggaaacatcc tcggacacaa
      781 gcttgagtac aacctcccag accagctcac cgaggagcag atcgccgagt tcaaggtaag
      841 tttaaacatg attttactaa ctaactaatc tgatttaaat tttcaggagg ccttctccct
      901 cttcgacaag gacggagacg gaaccatcac caccaaggag ctcggaaccg tcatgcgttc
      961 cctcggacag aacccaaccg aggccgagct ccaggacatg atcaacgagg tcgacgccga
     1021 cggagacgga accatcgact tcccagagtt cctcaccatg atggcccgta agatgaagta
     1081 ccgtgacacc gaggaggaga tccgtgaggc cttcggagtc ttcgacaagg acggaaacgg
     1141 atacatctcc gccgccgagc tccgtcacgt catgaccaac ctcggagaga agctcaccga
     1201 cgaggaggtc gacgagatga tccgtgaggc cgacatcgac ggagacggac aggtcaacta
     1261 cgaggagttc gtccagatga tgaccgccaa gggaggacgt ggaggatctg gcggcgcgcg
     1321 atccggagcc accaacttct ccctcctcaa gcaggccgga gacgtcgagg agaacccagg
     1381 accaggtacc atgaccgctc caaagaagaa acgcaaagta ccgagctcag aaaaaatgac
     1441 ggcacctaaa aagaaacgca aagtaccggt cgccgagaag acagccccaa aaaagaagcg
     1501 taaagttcca cgcgtcaaaa atgcgcctgt ctccaaggga gaggagctca tcaaggagaa
     1561 catgcacacc aagctctaca tggagggaac cgtcaacaac caccacttca agtgcaccca
     1621 cgaaggggag ggcaagccct acgagggcac ccagaccaac aggattaagg tggtggaggg
     1681 aggccccctg ccgttcgcat tcgacatcct ggccacctgc tttatgtacg ggagcaagac
     1741 cttcatcaac cacacccaag gaatcccaga cttcttcaag caatccttcg taagtttaaa
     1801 catgatttta ctaactaact aatctgattt aaattttcag cctgagggct tcacatggga
     1861 gagagtcacc acatacgaag acgggggcgt gcttaccgtt acccaggaca ccagcctcca
     1921 ggacggctgc ttgatctaca acgtcaagct cagaggggtg aacttcccat ccaacggccc
     1981 tgtgatgcag aagaaaacac tcggctggga ggccagtacc gagaccctgt accccgctga
     2041 cggcggcctg gaaggcagat gcgacatggc cctgaagctc gtgggcgggg gccacctgca
     2101 ctgcaacctg aagaccacat acagatccaa gaaacccgct aagaacctca agatgccagg
     2161 agtaagttta aacatatata tactaactaa ccctgattat ttaaattttc aggtctactt
     2221 tgtggaccgc agactggaaa gaatcaagga ggccgacaat gagacctacg tcgagcagca
     2281 cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca aacttaatgg
     2341 catggacgag ctgtacaagt aagctcagca ttcgtagaat tgtaagttta aactttctca
     2401 tactaactaa cgcgctctat ttaaattttc agaattccaa ctgagcgccg gtcgctacca
     2461 ttaccaactt gtctggtgtc aaaaataata ggggccgctg tcatcagagt aagtttaaac
     2521 tgagttctac taactaacga gtaatattta aattttcagc atctcgcgcc cgtgcctctg
     2581 acttctaagt ccaattactc ttcaacatcc ctacatgctc tttctccctg tgctcccacc
     2641 ccctattttt gttattatca aaaaacttct cttaatttct ttgtttttta gcttctttta
     2701 agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg taataaaaag
     2761 tcgaaaaaaa ttgtgctccc tccccccatt aataataatt ctatcccaaa atctacacaa
     2821 tgttctgtgt acacttctta tgttttttac ttctgataaa tttttttgaa acatcataga
     2881 aaaaaccgca cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt
     2941 atttcttcgc acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt
     3001 ggagtatttt tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt
     3061 gctttttggg gtttccccta ttgtttgtca agatttcgag gacggcgttt ttcttgctaa
     3121 aatcacaagt attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc
     3181 tcagtggaag gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt
     3241 gccttaaatg acagaataca ttcccaatat accaaacata actgtttcct actagtcggc
     3301 cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3361 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3421 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     3481 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3541 aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc
     3601 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3661 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3721 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3781 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3841 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3901 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3961 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4021 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4081 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4141 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4201 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4261 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4321 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4381 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4441 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4501 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4561 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4621 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4681 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4741 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4801 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4861 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4921 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4981 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5041 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5101 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5161 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5221 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5281 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5341 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5401 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5461 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5521 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5581 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5641 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5701 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5761 acaggaaaca gctatgacca tgattacgcc aagctgtaag tttaaacatg atcttactaa
     5821 ctaactattc tcatttaaat tttcagagct taaaaatggc tgaaatcact cacaacgatg
     5881 gatacgctaa caacttggaa atgaaataag cttgggctgc agcatgcggc cgctctaatc
     5941 actatcacgt gggatctgga taaattttct ccaaatttct cataaaaaat ggaaaaaaag
     6001 aaaaggaaat ccggtttgtg tatttccatt ttcaatttga tgcattaaac cagaacttcg
     6061 ttaatttccc ttctcttctc cctcaagcgt gcacactttt tttagatgag aagggctcta
     6121 tttcgattgg ttgtttttgt tttgaaccag attttttttt cacaaaaata aaatttggaa
     6181 ccttttgcca gtttaataaa gttttttgtt gggaatgttc ctaaaaattt ttaatttatc
     6241 taatataaaa cttgatagaa acattaatta acctgctttt ttgcatttat ttccaactga
     6301 attaaaataa atgtgaattt tttttactat gtaaataaat cgaaaattct ggttcctgat
     6361 tctcaagacc gcatctcaac ttttcagccg gtaatttgtc cgagaggagt ccaattttca
     6421 aatttttgat caaaatgtct actttgatcg acatcctcct tcttctgcaa ggtgtgtatc
     6481 atcgtcttga aatttcaaat tccctaaggg aacgaaaagc ttgtctcttt tggcagattc
     6541 gattcgattt cacctctttc tcccccctcc tgcctacaat gttgaaaggg tcccctcctt
     6601 gtgctccaaa ttgctcataa actcctccat actggacgct tctttttgcg gaaattggag
     6661 tcccagatat attggaagag ataaacggtt ttcgggtttg cagttgttgc cattttcact
     6721 gatttttatt caagtagtac tacttgaagt tttgttcctg agtaaaattt tataggttcc
     6781 atgatttatg aaattgttct atagtagcac caaacttttc agaacttttt tgtttttttt
     6841 ttcaatttta cagtaaaaag ttattctcaa atgttttctc aatttttgcc ccttattaac
     6901 tcacgtacgt atgtagttta cagagcgtat gtagatgtag ttccagtttt tggcacttat
     6961 aattttagaa gtcgaccgat aataaatgtt taggcatgcc aaatcatcat aagctactca
     7021 aaaaatagaa aatttttgat atgtagcaat tttctaaaac tttatctaaa aatctttttt
     7081 tccaaaaaat gcctttggtt aataaattgt aaatcaaaat tcagcaacaa aaaacctcac
     7141 tggaaaagtg ggagtaaaaa tacgaaataa aattttcgta actttttgtc aaaacagaat
     7201 cttaaatttc ccactatacc catttgtcct tcaaaaatgc catttaaagt aagcaaagag
     7261 gtatcataaa ttctcacaga acgtatgtag tttttgtgtc ctttacttct ccattgcctc
     7321 atattttcac ttttttcgtg gccgcagctc acagtgcaca catcacagaa tgttccagag
     7381 ccgaatgatt ttggatggaa tatcgagtga tgaatattca cccgggattg gccaaaggac
     7441 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg
     7501 ctagcgtcga cggatccaaa aaatgggatc tcaccaccac caccaccacg gaatggcctc
     7561 catgaccgga ggacagcaga tgggacgtga cctctacgac gacgacgaca aggacctcgc
     7621 cactatggtc gactcctccc gtcgtaagtg gaacaagacc ggacacgccg tccgtgccat
     7681 cggacgtctc tcctccctcg agaacgtcta catcaaggcc gacaagcaga agaa
//
