LOCUS       sequence-377522-        7974 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Pacr-2s-GCaMP6s::wCherry unc-54 3' UTR C.elegans A/B MN expression 
            of GCaMP6s RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7974)
  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 7974)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pJH3666
FEATURES             Location/Qualifiers
     source          1..7974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2019..2036
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(2076..2084,2085..2099)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2127..2183
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(5414..5436)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5536..5555
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5633..5651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5719..5823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5824..5892,5893..6684)
                     /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(6042..6061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6855..7443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7344..7363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7597..7614
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7731..7752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7767..7772,7773..7790,7791..7797)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7805..7821
                     /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     7810..7832
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7829..7845
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7829..7845
                     /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 gggattggcc
     1921 aaaggaccca aaggtatgtt tcgaatgata ctaacataac atagaacatt ttcaggagga
     1981 cccttggcta gcgtcgacgg atccaaaaaa tgggatctca ccaccaccac caccacggaa
     2041 tggcctccat gaccggagga cagcagatgg gacgtgacct ctacgacgac gacgacaagg
     2101 acctcgccac tatggtcgac tcctcccgtc gtaagtggaa caagaccgga cacgccgtcc
     2161 gtgccatcgg acgtctctcc tccctcgaga acgtctacat caaggccgac aagcagaaga
     2221 acggaatcaa ggtaagttta aacatgattt tactaactaa ctaatctgat ttaaattttc
     2281 aggccaactt ccacatccgt cacaacatcg aggacggagg agtccagctc gcctaccact
     2341 accagcagaa caccccaatc ggagacggac cagtcctcct cccagacaac cactacctct
     2401 ccgtccagtc caagctctcc aaggacccaa acgagaagcg tgaccacatg gtcctccttg
     2461 agttcgtcac cgccgccgga atcaccctcg gaatggacga gctctacaag ggaggaaccg
     2521 gaggttccat ggtctccaag ggagaggagc tcttcaccgg agtcgtccca atcctcgtcg
     2581 agctcgacgg agacgtcaac ggacacaagt tctccgtctc tggagaggta agtttaaaca
     2641 tgattttact aactaactaa tctgatttaa attttcaggg agagggagac gccacctacg
     2701 gaaagctcac cctcaagttc atctgcacca ccggaaagct cccagtcccg tggccgaccc
     2761 tcgtcaccac cctcacctac ggagtccagt gcttctcccg ttacccagac cacatgaagc
     2821 agcacgactt cttcaagtcc gccatgccag agggatacat ccaggagcgt accatcttct
     2881 tcaaggacga cggaaactac aagacccgtg ccgaggtcaa gttcgaggga gacaccctcg
     2941 tcaaccgtat cgagctcaag ggaatcgact tcaaggagga cggaaacatc ctcggacaca
     3001 agcttgagta caacctccca gaccagctca ccgaggagca gatcgccgag ttcaaggtaa
     3061 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcaggag gccttctccc
     3121 tcttcgacaa ggacggagac ggaaccatca ccaccaagga gctcggaacc gtcatgcgtt
     3181 ccctcggaca gaacccaacc gaggccgagc tccaggacat gatcaacgag gtcgacgccg
     3241 acggagacgg aaccatcgac ttcccagagt tcctcaccat gatggcccgt aagatgaagt
     3301 accgtgacac cgaggaggag atccgtgagg ccttcggagt cttcgacaag gacggaaacg
     3361 gatacatctc cgccgccgag ctccgtcacg tcatgaccaa cctcggagag aagctcaccg
     3421 acgaggaggt cgacgagatg atccgtgagg ccgacatcga cggagacgga caggtcaact
     3481 acgaggagtt cgtccagatg atgaccgcca agggaggacg tggaggatct ggcggcgcgc
     3541 caaaaatggt ctcaaagggt gaagaagata acatggcaat tattaaagag tttatgcgtt
     3601 tcaaggtgca tatggaggga tctgtcaatg ggcatgagtt tgaaattgaa ggtgaaggag
     3661 aaggccgacc atatgaggga acacaaaccg caaaactaaa ggtaagttta aacatatata
     3721 tactaactaa ccctgattat ttaaattttc aggtaactaa aggcggacca ttaccattcg
     3781 cctgggacat cctctctcca cagttcatgt atggaagtaa agcttatgtt aaacatccgg
     3841 cagatatacc agattatttg aaactttcat tcccggaggg ttttaagtgg gaacgcgtaa
     3901 tgaattttga agacggagga gttgttacag tgacgcaaga ctcaaggtaa gtttaaacag
     3961 ttcggtacta actaaccata catatttaaa ttttcagcct ccaagatgga gaatttattt
     4021 ataaagtcaa acttcgagga acgaatttcc cctcggatgg atctgttatg cagaagaaga
     4081 ctatgggatg ggaagcttca agtgaaagaa tgtaccctga agacggtgct cttaagggag
     4141 agattaaaca acgtcttaaa ttgaaagatg gaggacatta cgatgctgag gtaagtttaa
     4201 acatgatttt actaactaac taatctgatt taaattttca ggtgaagaca acttacaaag
     4261 ccaaaaaacc agttcagctg ccaggagcgt acaatgttaa tattaaactg gatatcacct
     4321 cccacaacga ggattacact atcgttgagc aatatgaaag agctgaaggg cggcactcga
     4381 caggtggcat ggatgaattg tataagtagg agctcccgag atcttcagcg ctcagcattc
     4441 gtagaattgt aagtttaaac tttctcatac taactaacgc gctctattta aattttcaga
     4501 attccaactg agcgccggtc gctaccatta ccaacttgtc tggtgtcaaa aataataggg
     4561 gccgctgtca tcagagtaag tttaaactga gttctactaa ctaacgagta atatttaaat
     4621 tttcagcatc tcgcgcccgt gcctctgact tctaagtcca attactcttc aacatcccta
     4681 catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa aacttctctt
     4741 aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg tgtagattca
     4801 aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc ccccattaat
     4861 aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt tttttacttc
     4921 tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt
     4981 acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc
     5041 aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa
     5101 gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg tttgtcaaga
     5161 tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa
     5221 gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga
     5281 aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc
     5341 aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg
     5401 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5461 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5521 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     5581 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     5641 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5821 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5881 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5941 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6001 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6061 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6121 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6181 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6241 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6301 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6361 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6421 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6481 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6541 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6601 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6661 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6721 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6781 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6841 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6901 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6961 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7021 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7081 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7141 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7201 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7261 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7321 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7381 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7441 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7501 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7561 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7621 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7681 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7741 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7801 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7861 ctgtaagttt aaacatgatc ttactaacta actattctca tttaaatttt cagagcttaa
     7921 aaatggctga aatcactcac aacgatggat acgctaacaa cttggaaatg aaat
//
