LOCUS       sequence-377517-        8685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pacr-2s-loxP::EBFP::loxP::Chrimson::wCherry unc-54 3' UTR C.elegans 
            A/B MN expression of Arch RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8685)
  AUTHORS   Lu Y, Ahamed T, Mulcahy B, Meng J, Witvliet D, Guan SA, Holmyard D, 
            Hung W, Wen Q, Chisholm AD, Samuel ADT, Zhen M
  TITLE     Extrasynaptic signaling enables an asymmetric juvenile motor circuit
            to produce symmetric undulation.
  JOURNAL   Curr Biol. 2022 Nov 7;32(21):4631-4644.e5. doi: 
            10.1016/j.cub.2022.09.002. Epub 2022 Sep 30.
  PUBMED    36182701
REFERENCE   2  (bases 1 to 8685)
  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"; doi: 
            "10.1016/j.cub.2022.09.002"; journalName: "Curr Biol"; date: 
            "2022-11-7- 7"; volume: "32"; issue: "21"; pages: "4631-4644.e5"
COMMENT     Sequence Label: pJH4140
FEATURES             Location/Qualifiers
     source          1..8685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1914..1947)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      2824..3147
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(3151..3184)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3762..3773
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             4419..5126
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPNYLKLSFPEGFKWERVMNFE
                     DGGVVTMTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4561..4579)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4829..4848
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(6125..6147)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6247..6266
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6344..6362)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6430..6534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6535..6603,6604..7395)
                     /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(6753..6772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7566..8154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8055..8074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8308..8325
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8442..8463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8478..8483,8484..8501,8502..8508)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8516..8532
                     /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     8521..8543
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8540..8556
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8540..8556
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcagcatgc ggccgctcac caccgactga ttttcatttc aaaattaatg
       61 cagatttcta cgtgttcttt ttcaaataaa tttatatatt ttttgttctc tcacaaaata
      121 aataatttcg agagttaagt gatcattctc agacatattc tataattgtc cgtcaggaaa
      181 cagatagagg aacctttgat aaataaatta taaaaggccg acagggaaat gcttctccct
      241 cgactctgtt agaagcgaag aggcaatttt taaaaatttc ccatttttat gcgtttttct
      301 gatttccaaa gtcacgcatc atttaccaac cgcaacgatt cataaggacg taattttgat
      361 tttctggacg atttttatat agttttgatg tgaaattcgc tttatttgtc tgaaagtgga
      421 atagatttag tgtggattta accgaaaaat gcagatcttt taaaaattta aaccagtgtt
      481 ttgttgcctt atttgcgaaa aaaaattgtg atatttcgag tattgctgca atttgattag
      541 taaaatgtta ggatataagt atttaaaaag tataattggt tgaattttct ttttcatcac
      601 cgacttccaa aaatatagag atactgagtt ttcaccattt tttgaatgta aataaaaaca
      661 ttttgaacac ttgtgctggg attcaatcca aatacattca ctgtcattta atattattat
      721 atatcaaaat aaaacgcata actttttctg ttttttttta aattttttca aaatgtaggt
      781 ttggtaagtt aaggtaaggt taggcaatgt aggtaaacac gagatgatac tccacgttga
      841 aaattgtttt cataaataca gtttagattg aactgaagga ttttatcgtc taatagaata
      901 taatttttta gttcaagatt taggatttgt tttcactttt ttttagattc aagacatata
      961 tctcgcaaaa tgagttttgt ctttgagcca tcttgtcatc atagcacatt gaattttgtt
     1021 gttcgcaaga cggaatatat gttcccagct gcaaattttg tgtcggatat ttcttcttta
     1081 gcgaaaggcg ttaaaaacca aaaatagtcg aagcgtttga tatgtgtccc gttggttttt
     1141 cgtgtccatc tattctgttt cgagtacaca tgttgcgacg cggcactgat gaaaaagtga
     1201 aagagctagt gtcacacaca aacacaaagt atgctcatgt gagtgtgtag atatgtgtag
     1261 atgtatgtcc tgaaggaata aaaagacttt ttgaagctac atttttaaac aaaaaaagac
     1321 agaaacatga agcacaactc tctattctgt cgaacggcgt gtgcaactca cgcacaccca
     1381 aaaatatgaa atataacatt cgcaccaaca aagcgcgaat atcaatgaat ctcgagggaa
     1441 cgatggctgc ctgccggcgc gtgttgctct tctgtcattg ggataactgg cgggtctgtt
     1501 gggtccgtcc tttagaccat ttgactacac tgccctgcat atgtgtgaaa cgtcgtcgtt
     1561 gctgttggga gctgtctgtg tgtatgtgtg tgtgtgtgag tatggagcag taaaaagctg
     1621 ccgatacgag aagcgccgcc ccggacggac ggcgatggcg catcgccaaa tgtgtcattt
     1681 tagttgtctt cacgaaccag agggcccacg acggacgggc ggccacgccc acctcaacca
     1741 gaatatcctt ccttctccca cccacccttg cggacgcgcg gatgcgctga acaaaaagac
     1801 gacgaagaag aagaagaaga agctgaacga agaaaaaagc atcaaaagaa gtgaagcggc
     1861 tctaccaccc cgaccacctt ccttcacagg aaggacgccg ttttcatccc gggataactt
     1921 cgtatagcat acattatacg aagttatatg gtctccaagg gagaggagct cttcaccgga
     1981 gtcgtcccaa tcctcgtcga gctcgacgga gacgtcaacg gacacaagtt ctccgtccgt
     2041 ggagagggag agggagacgc caccaacgga aagctcaccc tcaagttcat ctgcaccacc
     2101 ggaaagctcc cagtcccagt aagtttaaac atatatatac taactaaccc tgattattta
     2161 aattttcagt ggccaaccct cgtcaccacc ctctcccacg gagtccagtg cttcgcccgt
     2221 tacccagacc acatgaagca gcacgacttc ttcaagtccg ccatgccaga gggatacgtc
     2281 caggagcgta ccatcttctt caaggacgac ggaacctaca agagtaagtt taaacatgat
     2341 tttactaact aactaatctg atttaaattt tcagcccgtg ccgaggtcaa gttcgaggga
     2401 gacaccctcg tcaaccgtat cgagctcaag ggagtcgact tcaaggagga cggaaacatc
     2461 ctcggacaca agctcgagta caacttcaac tcccacaaca tctacatcat ggccgtcaag
     2521 cagaagaacg gaatcaaggt aagtttaaac atgattttac taactaacta atctgattta
     2581 aattttcagg tcaacttcaa gatccgtcac aacgtcgagg acggatccgt ccagctcgcc
     2641 gaccactacc agcagaacac cccaatcgga gacggaccag tcctcctccc agactcccac
     2701 tacctctcca cccagtccgt cctctccaag gacccaaacg agaagcgtga ccacatggtc
     2761 ctcctcgagt tccgtaccgc cgccggaatc accctcggaa tggacgagct ctacaagtga
     2821 taaatgcaaa atcctttcaa gcattccctt cttctctatc actcttcttt ctttttgtca
     2881 aaaaattctc tcgctaattt atttgctttt ttaatgttat tattttatga ctttttatag
     2941 tcactgaaaa gtttgcatct gagtgaagtg aatgctatca aaatgtgatt ctgtctgatg
     3001 tactttcaca atctctcttc aattccattt tgaagtgctt taaacccgaa aggttgagaa
     3061 aaatgcgagc gctcaaatat ttgtattgtg ttcgttgagt gacccaacaa aaagaggaaa
     3121 ctttattgtg ccgccaagaa aaaagtctca ataacttcgt atagcataca ttatacgaag
     3181 ttatgctagc gatccctatg gctgagctca tctcctccgc tacccgttcc ctcttcgctg
     3241 ctggaggaat caacccatgg ccgaacccat accaccacga ggacatggga tgcggaggaa
     3301 tgaccccaac cggagagtgc ttctccaccg agtggtggtg cgacccatcc tacggactct
     3361 ccgacgctgg atacggatac tgcttcgtcg aggctaccgg aggtaagttt aaacttggac
     3421 ttactaacta accatacata tttaaatttt caggatacct cgtcgtcgga gtcgagaaga
     3481 agcaggcttg gctccactcc cgtggaaccc caggagagaa gatcggagct caggtctgcc
     3541 agtggatcgc tttctccatc gctatcgctc tcctcacctt ctacggattc tccgcttgga
     3601 aggctacctg cggatgggag gaggtctacg tctgctgcgt cgaggtcctc ttcgtcaccc
     3661 tcgagatctt caaggagttc tcctccccag ctaccgtcta cctctccacc ggaaaccacg
     3721 cttactgcct ccgttacttc gagtggctcc tctcctgccc agtcatcctc atccgtctct
     3781 ccaacctctc cggactcaag gtaagtttaa acagttcggt actaactaac cctgattatt
     3841 taaattttca gaacgactac tccaagcgta ccatgggact catcgtctcc tgcgtcggaa
     3901 tgatcgtctt cggaatggct gctggactcg ctaccgactg gctcaagtgg ctcctctaca
     3961 tcgtctcctg catctacgga ggatacatgt acttccaggc tgctaagtgc tacgtcgagg
     4021 ctaaccactc cgtcccaaag ggacactgcc gtatggtcgt caagctcatg gcttacgctt
     4081 acttcgcttc ctggggttcc tacccaatcc tctgggctgt cggaccaggt aagtttaaac
     4141 atatatatac taactaacgg attatattta aattttcaga gggactcctc aagctctccc
     4201 catacgctaa ctccatcgga cactccatct gcgacatcat cgctaaggag ttctggacct
     4261 tcctcgctca ccacctccgt atcaagatcc acgagcacat cctcatccac ggagacatcc
     4321 gtaagaccac caagatggag atcggaggag aggaggtcga ggtcgaggag ttcgtcgagg
     4381 aggaggacga ggacaccgtc gcggtacccg cggccgccgt gagcaagggc gaggaggata
     4441 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     4501 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     4561 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     4621 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc aactacttga
     4681 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     4741 tggtgaccat gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     4801 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     4861 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     4921 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     4981 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     5041 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     5101 gcggcatgga cgagctgtac aagtaataga gatcttcagc gctcagcatt cgtagaattg
     5161 taagtttaaa ctttctcata ctaactaacg cgctctattt aaattttcag aattccaact
     5221 gagcgccggt cgctaccatt accaacttgt ctggtgtcaa aaataatagg ggccgctgtc
     5281 atcagagtaa gtttaaactg agttctacta actaacgagt aatatttaaa ttttcagcat
     5341 ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt
     5401 tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt
     5461 gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa
     5521 ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct
     5581 atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt
     5641 tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag
     5701 tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc
     5761 tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa
     5821 attaattttc ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga
     5881 cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag
     5941 aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt
     6001 agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac
     6061 tgtttcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     6121 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6181 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     6241 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6301 acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta
     6361 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6421 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6481 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6541 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6601 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6661 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6721 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6781 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6841 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6901 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6961 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7021 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7081 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7141 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7201 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7261 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7321 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7381 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7441 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7501 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7561 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7621 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7681 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7741 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7801 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7861 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7921 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7981 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8041 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8101 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8161 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8221 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8281 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     8341 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     8401 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     8461 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     8521 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctgtaagtt
     8581 taaacatgat cttactaact aactattctc atttaaattt tcagagctta aaaatggctg
     8641 aaatcactca caacgatgga tacgctaaca acttggaaat gaaat
//
