LOCUS       sequence-412784-        8495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pnpr-4 ins-22::GFP unc-54 3' UTR C.elegans AVA and other neurons 
            expression of ins-22 GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8495)
  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 8495)
  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: pJH4677
FEATURES             Location/Qualifiers
     source          1..8495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(4234..4236,4237..4749)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIE"
     primer_bind     complement(4276..4297)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4537..4556)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4884..4905
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5935..5957)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6057..6076
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6154..6172)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6240..6344
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6345..6413,6414..7205)
                     /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(6563..6582)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7376..7964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7865..7884
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8118..8135
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8252..8273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8288..8293,8294..8311,8312..8318)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8326..8342
                     /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     8331..8353
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8350..8366
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8350..8366
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcagcatgc ggccgcatca tgacatcagt attctgctat tcagctaaga
       61 ttcttgttgt cgtcatttga tgtatccgtt cgtcggtgca ggacttccac atggcgcagt
      121 cgaaaccatg ttaatctctg aagtcaaatg gtctaagaat tcacagtata gacggactgc
      181 aggtgtaaac ttatcctata tatcacccaa gacgtattac cttgtcagac tgcatttaca
      241 caagtaatcc cacttattat atgatgtcac gcgttataaa acggaagctt acttattgtg
      301 gaacaggtgg aatactcaaa aacattgaaa attcttacaa tttctaagta aaaaccttat
      361 gaatgttttg tacgtattat tattcccatt cttccattta gttgtcagtt tttcattgct
      421 cgggaagatt tgcataaata tcgccattga ccagtctact ccccagtttt ttgatcatga
      481 tttttcacaa caatgatcaa tcaaataaac atcatatcat cttatcccga ttaagcgccc
      541 tcgcaatttt attgcctcat cttcttaata atagaaacca tatgctcatt tttggcttag
      601 tttttgctca gttcaatttt ccttattcca tcgtaagcgt taagtctaga aggctttttg
      661 ttattcactt tttttctttg ctttttactt ttctctttct tcacgagaca aatattggta
      721 ttattgctca tctctctact aataaaactc aacaatttcg ttgaaacttg gtctctagag
      781 tttgatgagt tgtttgtgtt atttttgatc taaattttca attttgtgga agtaaggctt
      841 gcaagataga caaagacaac aagaaaataa acgaagggat tgaggaagaa taaactgaaa
      901 tgatcatggt taagaaaata ggaagaaaat agaagaagag cggccggctc actaggttaa
      961 attatttttg tgcttgttgt ggggagcgtt gagtaagacc tagagcaagg aacgcggtta
     1021 gcaatccctg ccgtttccaa atctccagag actatgcgaa attcagaaga cacgggacat
     1081 caagacagat aggtacggag taagaccttc aaattgtttc ttctcttctt gattactctc
     1141 ttgctcctct ctcgtccatt gtgctctttc ccacacactg agcaaaagca tcgtcattgt
     1201 ggaaagtgag cttgatgtgg ttggctcaaa catctgacca gctagacttg tcacttgtaa
     1261 gatatcacat cgcacttccc aattaatctt aaaatccgct tgactcattt atttttgaat
     1321 tcgtttttgt ttgaaattgt aaacttcgga atagttccag accgattcag aactagtatg
     1381 atcacctatg aaacgtgatt tctgaaatat tcttactttt ttctagacaa ctcaagatct
     1441 cgatcggcca aaattgtatt atcctcgtaa tcatcataac catttccgat attctgatca
     1501 caacggcttt ctttcgaatc tagtacgtct cacgtacaac tgcccacaaa cgatttccat
     1561 gaaactgata aataattaaa gactttggac ttaccaagag ttcaaacagg ttttatttcg
     1621 ttttctattt ttgaaaaaaa aatcattcca gttggacgac aaaaacagtt gtagaacttt
     1681 aataataata tttattcttg acacaaaaaa acttatcgta tttgtaaggg gaaaatagac
     1741 ccataattaa caacgtttct attagttatg cattttactg tactcatgat attactccaa
     1801 taacaaataa tttatctgaa atagcacttt tctccttctc cctgtgctcc aatacagtcc
     1861 agacacatat taattttcaa caccacatca cgcacatctt ccataggaaa ttaaaaaaaa
     1921 aatacattag acttcctatt catatcaaat atttcacatc ataatatttt tgtttgctca
     1981 tttttcaaca cttttctgat tcattttatt tttgtacgtt acgataccta catcagttat
     2041 ttttcacctg ccgtcactta ttttagatat tccttaattt tgactacgta ttactcattg
     2101 ttaacatttc tgatatttgt atactcaaag ccatttggaa agaatcagaa cagtgatttc
     2161 agttttgatg ctggccaact agttgaataa cgtgatgagg accattgagt aattagagaa
     2221 aacatgttta cgaatatagt cggctaaatt gagaaaaata cctaaataac tggcaataag
     2281 ctcagaactg aaaaacaaaa aattgctaac catcccgtga atttttgatg tgtgcctgtc
     2341 tatgccgtca tgttcatctc atgtgctttg ttttcaatcc ctggtctctt gtaatttttc
     2401 ttttaaaaag aagtctgcta ttcgtgacaa attaaaattg aaaattcata tatctttaga
     2461 gtatagccgt tttgagtcag caacaaacaa ttaggtgcca cttccgggac aggcacctct
     2521 gaaataaaag gaataagaaa atagtgagat attcccccct cactgatcaa acttcttttt
     2581 ttccctttac tacgtttacc tatattttta ttctgttagt ttggtcgaaa attggtggag
     2641 gatgtctaga atttgcagga tacaaattga gactaaagaa aattgaattt cttcaagtct
     2701 tctcgatttc ttgatttcta gtaaagcttc ctttgtaact actgtaatag ttagtgagaa
     2761 taattaaatt ttggcgcctg ccagtcattt ctagtgaaca aatataacat caatactttg
     2821 aaatctaatt ttctaagtag ctcgggtaat ttttttgttc acatcactcc ttacaacaca
     2881 actttcttca aaaatttcac gtctatattt agtttcaatt tttcaaaggt tgaaggcgtc
     2941 tagacttcga tgaataccaa aattaaagca atcatagttt tttaggtaaa gtttatgaat
     3001 ctgcaaagtc atactataaa actgatctta gaaatttttc ttgcagattt tatcttaata
     3061 tttcaaaaaa catcgttatg ttcccatttt gatttttggt gtaatggtcg tgtttctacg
     3121 aacaaaaacg gagacacccc caaaaatttt tgagatgctg tttttgaaaa taataaaggc
     3181 agctgtcagc tttattcatt tttctgaaaa acaattcact gtcattttga gctaatagct
     3241 gcaaaaccaa caaacgtaaa aactcagaaa gcattcaaaa atcgtaaaac ttgagttttt
     3301 ctcaagaaat acgattaaga ttactgtaga caaatttata gtaaattgaa ttaaagtgta
     3361 atttgataaa tgcatgactt tctcagattt attcattttc cacatataaa ctctaattga
     3421 atactaggaa ctgtgtatga gtatatcatt tgttgaaatg ctacattttt ctgttagtct
     3481 agatttttta acgagtttgt gtcttggtaa ttgtcgtgtg tattcattgt tcacagcttc
     3541 taataggaaa gggttatttt caaaaaacgt tctgtacttc gctggtagga gtactgtagt
     3601 tttactattt tttttatata acagaactat ttgtttctgc tttgaaagca taattcgaat
     3661 aattacgtag gattcaattt ttcttgatac tcattttctt ttttgtctaa ccatttcatt
     3721 ctgacacata aattaccaat aacagattga ttgtctgtac tttatgttat tttcctttca
     3781 atatttttcc tctctcttat ttctaaaagc ggaaatgaaa aacgactctg agaattaatt
     3841 ttcttcattt cttctgtagg cgtaactgca ctttctggga tttctttttg ctgtcataat
     3901 acgggagtac ccgggatgca cactacaact attctcatat gctttttcat ccttcttgtt
     3961 caagtctcca caatggatgc tcacactgac aaatacgtca gaactctgtg tggaaaaact
     4021 gcaatcagaa atattgccaa cctttgcccg ccaaagccag aaatgaaggg tatctgttct
     4081 accggaggtg agataaactt aaactcaaaa gccttgtttg caaataatta tttttctaaa
     4141 ttttcagagt atccaagcat caccgaatac tgttccatgg gattttcaga ctctcagatc
     4201 aagtttatgt gctgtgataa ccaatgggta cctatggtga agggcgagga gctgttcacc
     4261 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     4321 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     4381 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     4441 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     4501 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     4561 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     4621 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     4681 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     4741 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     4801 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     4861 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4921 actctcggca tggatgagct gtacaagtag gctcagcatt cgtagaattg taagtttaaa
     4981 ctttctcata ctaactaacg cgctctattt aaattttcag aattccaact gagcgccggt
     5041 cgctaccatt accaacttgt ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa
     5101 gtttaaactg agttctacta actaacgagt aatatttaaa ttttcagcat ctcgcgcccg
     5161 tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt tctccctgtg
     5221 ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc
     5281 ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta
     5341 ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat
     5401 ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac
     5461 atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg
     5521 caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa
     5581 aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc
     5641 ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt
     5701 cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg
     5761 tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg
     5821 gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac tgtttcctac
     5881 tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5941 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6001 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6061 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6121 aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta tttttatagg
     6181 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     6241 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6301 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6361 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6421 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6481 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6541 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6601 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6661 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6721 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6781 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6841 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6901 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6961 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7021 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7081 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7141 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7201 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7261 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7321 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7381 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7441 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7501 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7561 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7621 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7681 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7741 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7801 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7861 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7921 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7981 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8041 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8101 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     8161 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     8221 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     8281 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     8341 aatttcacac aggaaacagc tatgaccatg attacgccaa gctgtaagtt taaacatgat
     8401 cttactaact aactattctc atttaaattt tcagagctta aaaatggctg aaatcactca
     8461 caacgatgga tacgctaaca acttggaaat gaaat
//
