LOCUS       sequence-413947-        9231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pflp-18 LoxP EBFP (stop) LoxP twk-40(gf) GFP unc-54 3' UTR C.elegans
            AVA and other neurons expression of twk-40(gf) GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9231)
  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 9231)
  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: pJH4316
FEATURES             Location/Qualifiers
     source          1..9231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(429..440)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1558..1591)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      2468..2791
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2795..2828)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4908..4936)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5684..5704
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6671..6693)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6793..6812
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6890..6908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6976..7080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7081..7149,7150..7941)
                     /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(7299..7318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8112..8700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8601..8620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8854..8871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8988..9009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9024..9029,9030..9047,9048..9054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9062..9078
                     /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     9067..9089
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9086..9102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9086..9102
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcagcatgc agcttagccg gaatagggtc acgaaaacat gaaaataaag
       61 cttgaaccta ttgttatgat tttcaatgcc accttttgca gtttaatata cactattact
      121 acttctcagt tcaagttcac ttctgagacg gaacctttat tttctaactt tctttaatct
      181 tactttttat gagccgaacg ttctattttg taaaatgatc agatacaagc ccaaagctaa
      241 ttttgccaat aagtgttagt agatcaattg aattacaata taatgacctg ggcggaatag
      301 atttttgcac tgatttttct ccaagcttac tctatgaata ttcatagatc atcttcaaag
      361 gtagcgtgtg ttgtagttgt gttttggcgg gttcgtgtta tttgactcag ctcaagtaga
      421 tgttgtagtc ttccctcaat caacgcgtgg gcttgaccct ctttaaattg agtggaaacc
      481 agccaatcta tttgaaatct acccccgaaa tgagattcga cgatgacacg acgtgtgtgt
      541 tacgtgcgct gagaatttag gaaaaaaaag ataatcttct tttcgttagc tgcgccacaa
      601 cctgtgcaga taaactccgc acaattgaag ttcttactgg tcccacccga tttattcaat
      661 tgtattgtgt ctttttttct tatttctcta caactctaac tttcttcaac tattctctac
      721 atcagtgctt ttttccacca tgaaaaaaga aatgacgtcg ttgtttctta ggaggggcgg
      781 agcctcaact caatggtccg ttcgaataga tcccacccct catataaata ggactgaaac
      841 caaagaagtt tttaaatttt cagcttaccg tgtttctcaa ttttcaaaat agtctttttt
      901 gtgtctgaac gggctgatca attatgtgtt tttttgaatg aaaaatcttc ctcccaagcc
      961 ttaaaattcc ttccaaagat cgaaagttat gtctattcaa gttacttctt cttagatcaa
     1021 atattaaaca aaaaaatacg ccatcattct ctcacattta gtaattcaaa atttcgattt
     1081 gttttcgaac ttttgaattt cccgcctagg aagtgacatg agtgaaatct cattagataa
     1141 atgtggcact caccgatgac actattttca aataaggggg gggggtgttt cgaatacctt
     1201 gtacacgcgt ttccgcaaaa ccacacaccc cgagttcctc agcttcttcc cctcgaccca
     1261 tgagccacgt caagatgtca gcgtgtcact tttcattgct tcaccttcta cggtaacgag
     1321 aaaaggcggg gaatccagga tctggtgtga cttcttcctt cttcgtatct tcctaaaaat
     1381 gttcgtttac tttttagcga gtgtcatttg atgggatgtc gttcttgagg aaagaggcta
     1441 gaagcttaat actctcatat ttcatattat tggcatcaag acgtttatta tttaactaca
     1501 ataacgattg ctttaacaac taaaaataat aatttcaggg ttagacggat ccccgggata
     1561 acttcgtata gcatacatta tacgaagtta tatggtctcc aagggagagg agctcttcac
     1621 cggagtcgtc ccaatcctcg tcgagctcga cggagacgtc aacggacaca agttctccgt
     1681 ccgtggagag ggagagggag acgccaccaa cggaaagctc accctcaagt tcatctgcac
     1741 caccggaaag ctcccagtcc cagtaagttt aaacatatat atactaacta accctgatta
     1801 tttaaatttt cagtggccaa ccctcgtcac caccctctcc cacggagtcc agtgcttcgc
     1861 ccgttaccca gaccacatga agcagcacga cttcttcaag tccgccatgc cagagggata
     1921 cgtccaggag cgtaccatct tcttcaagga cgacggaacc tacaagagta agtttaaaca
     1981 tgattttact aactaactaa tctgatttaa attttcagcc cgtgccgagg tcaagttcga
     2041 gggagacacc ctcgtcaacc gtatcgagct caagggagtc gacttcaagg aggacggaaa
     2101 catcctcgga cacaagctcg agtacaactt caactcccac aacatctaca tcatggccgt
     2161 caagcagaag aacggaatca aggtaagttt aaacatgatt ttactaacta actaatctga
     2221 tttaaatttt caggtcaact tcaagatccg tcacaacgtc gaggacggat ccgtccagct
     2281 cgccgaccac taccagcaga acaccccaat cggagacgga ccagtcctcc tcccagactc
     2341 ccactacctc tccacccagt ccgtcctctc caaggaccca aacgagaagc gtgaccacat
     2401 ggtcctcctc gagttccgta ccgccgccgg aatcaccctc ggaatggacg agctctacaa
     2461 gtgataaatg caaaatcctt tcaagcattc ccttcttctc tatcactctt ctttcttttt
     2521 gtcaaaaaat tctctcgcta atttatttgc ttttttaatg ttattatttt atgacttttt
     2581 atagtcactg aaaagtttgc atctgagtga agtgaatgct atcaaaatgt gattctgtct
     2641 gatgtacttt cacaatctct cttcaattcc attttgaagt gctttaaacc cgaaaggttg
     2701 agaaaaatgc gagcgctcaa atatttgtat tgtgttcgtt gagtgaccca acaaaaagag
     2761 gaaactttat tgtgccgcca agaaaaaagt ctcaataact tcgtatagca tacattatac
     2821 gaagttatgc tagcatggca acatggaaaa cctacgcccg gataatcctc gctcacgtca
     2881 gtcttattgt cttgtcggtt gtctatgttg gatttggagc gtttctcttc tatcagttgg
     2941 agcaaccgaa tgaagttgaa gttcgagcta ggaatatcga aaggttcaat attcacaaac
     3001 gacaaatgat tgaacattta tgggagatga gggaaagcgg tattggacaa cacgtggtgg
     3061 aggatctggc tgtaaaatac gtagacaacg tcactcgcat tcttttcgag gcatttgaca
     3121 cccattgtat aggagctaag catctgagac ccggcggaga ggatgaagac gtgagttgga
     3181 gtctgaaatg tttatctgaa attgaatttg cactgtgttt aagtttcaca aactcaattt
     3241 ttaaatttca gtacaattgg acctacatga cagctttatt tttcactaca actctcctca
     3301 cgactattgg ttacggtaac ttgaccccag taaccggaag aggaaaattg ctctgcatcc
     3361 tatacgcact tttcggtgtc ccactgatca atataactgt tgccgatatc ggaaagtttt
     3421 tatctgaaaa tattgttcaa ttgtatactt ggtgagacac ttttttttga aaactttttt
     3481 aaaaatccga aattcaggta ccgaaagctt cgggaaaagt gctcaaaaca aaaatacagt
     3541 gtcatttcca gtaaggacga taagaataag gaaggggacc tgaatctggt aagagggtct
     3601 aaaaatggtc tagtagtggt taaaatgggc aaaaacaggc aaacattttt tgaaattaat
     3661 aagtacattt cgagtatact tttgaaatca gccaatatcg ggtacatttt taaaattcgg
     3721 aaaaaaaaac gagcgataaa accgcccgct aatcaccggc aacttgccca tctttcagct
     3781 tctacaaaaa attcaatttc tccagtaaac caaaaaaaaa acttgaaaaa aaaacaaaac
     3841 attttgtttt tttttctttt ttggtctgaa caaaaaacta aaaaaaaaca tccaaaaaat
     3901 tcatttttgc caaaaacacc aaaataacca aaacaatcga ccacttcacc ctcgaacact
     3961 ttccaattag aactttttaa ggatcatttg gaaaactaca tctcgattcc tatattccta
     4021 attgtggcga tactactcag ctacataaca tttggagcag ttgttctgag tatgtgggaa
     4081 ggatgggatt tcttctctgg attctacttt agctttatta caatgacgac tgtcggattt
     4141 ggtgatatag taccattgaa acgggaatat tatattttag acttatgcta tattattatt
     4201 ggattatcga ttacgtaagt ttgtgatttt ggctattagt gcctcgagtc tcaaaaaaac
     4261 tgtagtttgt agttgacaaa tttcaaatcg ttgaaaattt ttgccagttg cgttatttta
     4321 tataggtagt accaacaaaa aatttccagc acgatgtgta tcgatttggt tggaattcag
     4381 tatattcgga aaatgcatta cttcggaaga gctattaagg taagaagcta aatttttagt
     4441 cctactaaaa ttttctagga cgcccgattt gctttggtaa acgtcggcgg gaagatggtt
     4501 cacgtgccgg atttaatgag atatgcgagt gttctacaac aaaagtatgg ccaaaagaag
     4561 acccatgaca caataattgt caagggagct tatgctccaa aggatctgag caaaatacgg
     4621 tgagaggact cggttttttt cttaataaaa agaaaaatca atttaaattt ttgtatatct
     4681 agattcgtat aacagttcag ttacaaatta aatatttcaa atattcattt ggtatttact
     4741 ctgataagtt ttgatatctg aaatattttg aaaactcgta aatttccaga tttatcgatt
     4801 ttggagcatt ggcctcaatt gaaagttggc aatcgctatg gtcaattctc actggaagaa
     4861 ctcaagaagt tcatgttatg agtaaaggag aagaactttt cactggagtt gtcccaattc
     4921 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     4981 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg
     5041 ttccatgggt aagtttaaac atatatatac taactaaccc tgattattta aattttcagc
     5101 caacacttgt cactactttc tgttatggtg ttcaatgctt ctcgagatac ccagatcata
     5161 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     5221 tatttttcaa agatgacggg aactacaaga cacgtaagtt taaacagttc ggtactaact
     5281 aaccatacat atttaaattt tcaggtgctg aagtcaagtt tgaaggtgat acccttgtta
     5341 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
     5401 tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa
     5461 tcaaagttgt aagtttaaac atgattttac taactaacta atctgattta aattttcaga
     5521 acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac
     5581 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac
     5641 aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg
     5701 taacagctgc tgggattaca catggcatgg atgaactata caaatagcat tcgtagaatt
     5761 ccaactgagc gccggtcgct accattacca acttgtctgg tgtcaaaaat aataggggcc
     5821 gctgtcatca gagtaagttt aaactgagtt ctactaacta acgagtaata tttaaatttt
     5881 cagcatctcg cgcccgtgcc tctgacttct aagtccaatt actcttcaac atccctacat
     5941 gctctttctc cctgtgctcc caccccctat ttttgttatt atcaaaaaac ttctcttaat
     6001 ttctttgttt tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa
     6061 atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat
     6121 aattctatcc caaaatctac acaatgttct gtgtacactt cttatgtttt ttacttctga
     6181 taaatttttt tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg
     6241 tttcagttta tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa
     6301 tcatgctcat cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt
     6361 tatgaaatta attttcctgc ttttgctttt tggggtttcc cctattgttt gtcaagattt
     6421 cgaggacggc gtttttcttg ctaaaatcac aagtattgat gagcacgatg caagaaagat
     6481 cggaagaagg tttgggtttg aggctcagtg gaaggtgagt agaagttgat aatttgaaag
     6541 tggagtagtg tctatggggt ttttgcctta aatgacagaa tacattccca atataccaaa
     6601 cataactgtt tcctactagt cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg
     6661 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     6721 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     6781 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     6841 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg cctcgtgata
     6901 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     6961 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7021 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7081 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7141 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     7201 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     7261 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7321 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7381 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7441 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     7501 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7561 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7621 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7681 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7741 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7801 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7861 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7921 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7981 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     8041 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     8101 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     8161 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     8221 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     8281 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8341 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8401 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     8461 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8521 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8581 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8641 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8701 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8761 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8821 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8881 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     8941 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     9001 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     9061 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctg
     9121 taagtttaaa catgatctta ctaactaact attctcattt aaattttcag agcttaaaaa
     9181 tggctgaaat cactcacaac gatggatacg ctaacaactt ggaaatgaaa t
//
