LOCUS       sequence-408463-        5614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptwk-40s EGFP unc-54 3' UTR C.elegans AVA and other neurons 
            expression of EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5614)
  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 5614)
  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: pJH4693
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1519..1521,1522..2034)
                     /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(1561..1582)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1822..1841)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2169..2190
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3054..3076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3176..3195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3273..3291)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3359..3463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3464..3532,3533..4324)
                     /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(3682..3701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4495..5083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4984..5003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5237..5254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5371..5392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5407..5412,5413..5430,5431..5437)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5445..5461
                     /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     5450..5472
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5469..5485
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5469..5485
                     /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 tcacccctat ggtgaagggc gaggagctgt tcaccggggt ggtgcccatc
     1561 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1621 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1681 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1741 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1801 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1861 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1921 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1981 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2041 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2101 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     2161 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggat
     2221 gagctgtaca agtaggagct ccgcatcggc cgctgtcatc agatcgccat ctcgcgcccg
     2281 tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt tctccctgtg
     2341 ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc
     2401 ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta
     2461 ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat
     2521 ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac
     2581 atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg
     2641 caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa
     2701 aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc
     2761 ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt
     2821 cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg
     2881 tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg
     2941 gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac tgtttctact
     3001 agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     3061 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     3121 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     3181 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     3241 aggagaaaat accgcatcag gcggccttaa gggcctcgtg atacgcctat ttttataggt
     3301 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3361 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3421 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3841 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4021 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4321 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4621 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5281 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5341 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5401 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5461 atttcacaca ggaaacagct atgaccatga ttacgccaag ctgtaagttt aaacatgatc
     5521 ttactaacta actattctca tttaaatttt cagagcttaa aaatggctga aatcactcac
     5581 aacgatggat acgctaacaa cttggaaatg aaat
//
