LOCUS       sequence-413951-        8419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pflp-18 LoxP EBFP LoxP TeTx mCherry unc-54 3' UTR C.elegans AVA and 
            other neurons expression of TeTx RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8419)
  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 8419)
  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: pJH4508
FEATURES             Location/Qualifiers
     source          1..8419
                     /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)."
     CDS             4233..4943
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4378..4396)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4646..4665
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5859..5881)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5981..6000
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6078..6096)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6164..6268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6269..6337,6338..7129)
                     /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(6487..6506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7300..7888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7789..7808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8042..8059
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8176..8197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8212..8217,8218..8235,8236..8242)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8250..8266
                     /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     8255..8277
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8274..8290
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8274..8290
                     /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 tagcatgccg atcaccatca acaacttccg ttactccgac ccggttaaca
     2881 acgacaccat catcatgatg gaaccgccgt actgcaaagg tctagacatc tactacaaag
     2941 ccttcaaaat caccgaccgt atctggatcg ttccggaacg ttacgaattt ggtactaaac
     3001 cggaagactt caacccgccg tcctccctga tcgaaggtgc ttccgaatac tacgacccga
     3061 actacctgcg taccgactcc gacaaagacc gtttcctgca gaccatggtt aaactgttca
     3121 accgtatcaa aaacaacgtt gctggtgaag ctctgctgga caaaatcatc aacgctatcc
     3181 cgtacctggg taactcctac tccctgctgg acaaattcga caccaactcc aactccgttt
     3241 ccttcaacct gctggaacag gatccgtccg gtgctaccac caaatccgct atgctgacca
     3301 acctgatcat cttcggtccg ggtccggttc tgaacaaaaa cgaagttcgt ggtatcgttc
     3361 tgcgtgttga caacaaaaac tacttcccgt gccgtgacgg tttcggttcc atcatgcaga
     3421 tggctttctg cccggaatac gtaccgacct tcgacaacgt tatcgaaaac atcacctccc
     3481 tgaccatcgg taaatccaaa tacttccagg acccggctct gctgctgatg cacgaactga
     3541 tccacgttct gcacggtctg tacggcatgc aggtttcctc ccacgaaatc atcccgtcca
     3601 aacaggaaat ctacatgcag cacacctacc cgatctccgc tgaagagctc ttcaccttcg
     3661 gtggtcagga cgctaacctg atctccatcg atatcaaaaa cgacctgtac gaaaaaaccc
     3721 tgaacgacta caaagctatc gctaacaaac tgtcccaggt tacctcctgc aacgacccga
     3781 acatcgacat cgactcctac aaacagatct atcagcagaa gtatcagttt gataaggatt
     3841 ccaacggcca gtatatagtg aacgaggata agtttcagat actgtataac tccataatgt
     3901 atggctttac tgagatagag ctgggcaaga agtttaacat aaagactcgc ttaagctatt
     3961 tttccatgaa ccacgatcct gtgaagatac ctaacctgct ggatgatact atatataacg
     4021 atactgaggg ctttaacata gagtccaagg atctgaagtc cgagtataag ggccagaaca
     4081 tgagggtgaa cactaacgcc tttaggaacg tcgacggttc cggtctggtt tccaaactga
     4141 tcggtctgtg caaaaaaatc atcccgccga ccaacatccg tgaaaacctg tacaaccgta
     4201 ccgctagggt accggtagaa aaaatgctcg agatggtgag caagggcgag gaggataaca
     4261 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     4321 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     4381 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     4441 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     4501 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     4561 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     4621 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     4681 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     4741 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     4801 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     4861 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     4921 gcatggacga gctgtacaag taagaattcc aactgagcgc cggtcgctac cattaccaac
     4981 ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa actgagttct
     5041 actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc tgacttctaa
     5101 gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt
     5161 ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt taagtcacct
     5221 ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa
     5281 aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt
     5341 gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata gaaaaaaccg
     5401 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     5461 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     5521 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     5581 gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct aaaatcacaa
     5641 gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga
     5701 aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa
     5761 tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg gccgtacggg
     5821 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5881 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5941 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6001 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6061 atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6121 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6181 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6241 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6301 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6361 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6421 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6481 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6541 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6601 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6661 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6721 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6781 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6841 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6901 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6961 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7021 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7081 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7141 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7201 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7261 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7321 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7381 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7441 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7501 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7561 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7621 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7681 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7741 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7801 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7861 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7921 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7981 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     8041 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     8101 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     8161 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     8221 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     8281 cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat
     8341 tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct
     8401 aacaacttgg aaatgaaat
//
