LOCUS       sequence-355747-        8625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Punc-4::GCaMP6::RFP unc-54 3' UTR C.elegans A-MNs and others 
            expression of GCaMP6::RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8625)
  AUTHORS   Gao S, Guan SA, Fouad AD, Meng J, Kawano T, Huang YC, Li Y, Alcaire 
            S, Hung W, Lu Y, Qi YB, Jin Y, Alkema M, Fang-Yen C, Zhen M
  TITLE     Excitatory motor neurons are local oscillators for backward 
            locomotion.
  JOURNAL   Elife. 2018 Jan 23;7. pii: 29915. doi: 10.7554/eLife.29915.
  PUBMED    29360035
REFERENCE   2  (bases 1 to 8625)
  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: "Elife."; 
            date: "2018-01-23"; volume: "7. pii: 29915. doi"; pages: " 
            10.7554/eLife.29915"
COMMENT     Sequence Label: pJH3137
FEATURES             Location/Qualifiers
     source          1..8625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     368..385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    502..523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(538..543,544..561,562..568)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    576..592
                     /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     581..603
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     600..616
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     600..616
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3415..3432
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(3472..3480,3481..3495)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             3523..3579
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(6810..6832)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6932..6951
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7029..7047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7115..7219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7220..7288,7289..8080)
                     /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(7438..7457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8251..8625,1..214)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
       61 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      121 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      181 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      241 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      301 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      361 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
      421 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      481 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      541 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      601 aggaaacagc tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact
      661 aactattctc atttaaattt tcagagctta aaaatggctg aaatcactca caacgatgga
      721 tacgctaaca acttggaaat gaaataagct tgggctgcag catgcggccg cttccccaaa
      781 ttggaacagt gaaatactca atcaatcact ctggcggttt tctccacaaa gaacacgaaa
      841 ttggaaaagg gaaaagattg atttgtttga aggcgtgagg aaatcgaaaa aaaaatgttt
      901 caaaacaaat gaaatatttt aatgaataaa cccaggaatg ccaccttttg gtgcccgcct
      961 acaaaaaagt caactgttct taaactccta catagttgcc ctctctaacg aatgttgctt
     1021 tttttaaatt ttcattgaat ttgattatcg tctgtatgcg cctactctat cagtggaaaa
     1081 catgcccatt tcccacaaat ttactcgttt ggagctcaaa atgttaaatg gggcatcatg
     1141 gaattggcat ctcacagaaa gtggcggatg agttatatgt aataataatg gtggctgaag
     1201 tggaaaccgc aaaaaaagga tgtgtatttg agattttgta acggaatttg aatcagatac
     1261 acaaaatgag agaaatttat agattcttta aaatccacct gaagtcggca aatttttgaa
     1321 aggtctgttc gttcagtttt tcataagttt tatgaatttt gatttactac tgaagttcga
     1381 gaaaagccct ttaaccatac caataagaaa ttatgttaac tttacaagaa tattaacaaa
     1441 aaattattaa aaaactgtgt tcccctggca ttatgatatt tccaccgtca gttctctatt
     1501 aaaacttcaa ctctacacag tttcaactct caaatgagct gtccatcttt tgccttcagg
     1561 aaattatgct aatctgctga tttcgcgcac tttcaccaca ctatttccaa atagaaggag
     1621 gggtggtgca acacacgaga cactgcaaaa aggttcagtg aagatttatc tacttgtgag
     1681 atgagagcag gaggggtggg agaagtgtga tgattgacgg gatgacaaga acagtgatgt
     1741 ggtgtcgaaa tggaagatgc gctaatttgt ggggttttat gaaatttgaa taattacatt
     1801 tataacatct actagttagg cgtctgtgta ggtattgcct gcctgcctac aaggcatatg
     1861 tacgaggcag gttaaacctc ctagaaagct acggtacttt caaaccacat aacatttata
     1921 ctggtgtctt ttcgcgtctt cttcttgaac atgtagaccc tcaaaaagct aagttttaca
     1981 ctgtgtagtt tgctttgaaa tctacaaaat tggaggcacg ccttctggac tttgccgaca
     2041 cgcatacttt ctacctacca actgcctact tccacagtct agcgtgaact ccaataccat
     2101 tttaaaaatt gttttatctc aaattttcaa tttttaaatc caatttccca gttctaaaaa
     2161 aaaaattttt tttttaattt aaaaactctc atccttttta atgctcgttc ccattcggca
     2221 agacgtgtac caaacacaaa acaaattgac tggtgatcca tctcgaattt ctcgtttcaa
     2281 attagtcgtt gctatatata ttgtcttatg tgtggaaaac gaataagaaa aattagagaa
     2341 agatgagaga gcaggggcag aatgaaaatt tgcatggaaa ttgaagaaga tcactgcttt
     2401 ttccgatgaa tgatgactaa atagaatgag ttgagaggaa aataatcata attctaattt
     2461 taattagaaa tatgatatgg aaagtaagca aaaccacaag gcttttgctc tgaacttatt
     2521 tccgaaaaaa aaaagtgaaa ctgaaagtta gaattttcaa aataatattg cgagagtcaa
     2581 ccttcagaaa aattggcgag aaatcaaatt tgaattctct gaaaatttag ttaaaataca
     2641 tatgttatcc ttctttgttt tttggcctta gcctgccttc aaggcgacct gagcctatgc
     2701 gggtgcctaa taccgaaaca cacataattt ttttttttaa tctttaaata ctgtaacgag
     2761 tcgcgagatc cagcgagacc catagaatat gttgagcgct gcaaacaatc gtataactct
     2821 tgttcaaatt tttccactct gtatttttgc aagcagttca acaaatggtc atcattttta
     2881 taacccatat ctatattcga atctgacata acaaataata ctaagcacat cctcccacat
     2941 ctcttttcat tcaagttgcg caattcccct aataactatg gatggagggg ggaccgagcc
     3001 caaagattat tcaaaacgat aacgactttt tcatgcggcc aacgaaaaat aaaacttttc
     3061 tcatattctc ctatttccca tttcaagaat tacatcccca tgggacgttg aaatcacatc
     3121 atattagtat atgttaatag ttcttttcac tggaatatcg ccacgccccc cttccggtgg
     3181 ctcattggtt tcgagaagaa ggcggagccg aagtgggttc ttctttcttt ttttgtggta
     3241 gaagtctagc aggattaagc tactattcca ttagaggatc ttcttcttcc aaaaagtgaa
     3301 aatcccggga ttggccaaag gacccaaagg tatgtttcga atgatactaa cataacatag
     3361 aacattttca ggaggaccct tggctagcgt cgacggatcc aaaaaatggg atctcaccac
     3421 caccaccacc acggaatggc ctccatgacc ggaggacagc agatgggacg tgacctctac
     3481 gacgacgacg acaaggacct cgccactatg gtcgactcct cccgtcgtaa gtggaacaag
     3541 accggacacg ccgtccgtgc catcggacgt ctctcctccc tcgagaacgt ctacatcaag
     3601 gccgacaagc agaagaacgg aatcaaggta agtttaaaca tgattttact aactaactaa
     3661 tctgatttaa attttcaggc caacttccac atccgtcaca acatcgagga cggaggagtc
     3721 cagctcgcct accactacca gcagaacacc ccaatcggag acggaccagt cctcctccca
     3781 gacaaccact acctctccgt ccagtccaag ctctccaagg acccaaacga gaagcgtgac
     3841 cacatggtcc tccttgagtt cgtcaccgcc gccggaatca ccctcggaat ggacgagctc
     3901 tacaagggag gaaccggagg ttccatggtc tccaagggag aggagctctt caccggagtc
     3961 gtcccaatcc tcgtcgagct cgacggagac gtcaacggac acaagttctc cgtctctgga
     4021 gaggtaagtt taaacatgat tttactaact aactaatctg atttaaattt tcagggagag
     4081 ggagacgcca cctacggaaa gctcaccctc aagttcatct gcaccaccgg aaagctccca
     4141 gtcccgtggc cgaccctcgt caccaccctc acctacggag tccagtgctt ctcccgttac
     4201 ccagaccaca tgaagcagca cgacttcttc aagtccgcca tgccagaggg atacatccag
     4261 gagcgtacca tcttcttcaa ggacgacgga aactacaaga cccgtgccga ggtcaagttc
     4321 gagggagaca ccctcgtcaa ccgtatcgag ctcaagggaa tcgacttcaa ggaggacgga
     4381 aacatcctcg gacacaagct tgagtacaac ctcccagacc agctcaccga ggagcagatc
     4441 gccgagttca aggtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt
     4501 caggaggcct tctccctctt cgacaaggac ggagacggaa ccatcaccac caaggagctc
     4561 ggaaccgtca tgcgttccct cggacagaac ccaaccgagg ccgagctcca ggacatgatc
     4621 aacgaggtcg acgccgacgg agacggaacc atcgacttcc cagagttcct caccatgatg
     4681 gcccgtaaga tgaagtaccg tgacaccgag gaggagatcc gtgaggcctt cggagtcttc
     4741 gacaaggacg gaaacggata catctccgcc gccgagctcc gtcacgtcat gaccaacctc
     4801 ggagagaagc tcaccgacga ggaggtcgac gagatgatcc gtgaggccga catcgacgga
     4861 gacggacagg tcaactacga ggagttcgtc cagatgatga ccgccaaggg aggacgtgga
     4921 ggatctggcg gcgcgccaaa aatggtctca aagggtgaag aagataacat ggcaattatt
     4981 aaagagttta tgcgtttcaa ggtgcatatg gagggatctg tcaatgggca tgagtttgaa
     5041 attgaaggtg aaggagaagg ccgaccatat gagggaacac aaaccgcaaa actaaaggta
     5101 agtttaaaca tatatatact aactaaccct gattatttaa attttcaggt aactaaaggc
     5161 ggaccattac cattcgcctg ggacatcctc tctccacagt tcatgtatgg aagtaaagct
     5221 tatgttaaac atccggcaga tataccagat tatttgaaac tttcattccc ggagggtttt
     5281 aagtgggaac gcgtaatgaa ttttgaagac ggaggagttg ttacagtgac gcaagactca
     5341 aggtaagttt aaacagttcg gtactaacta accatacata tttaaatttt cagcctccaa
     5401 gatggagaat ttatttataa agtcaaactt cgaggaacga atttcccctc ggatggatct
     5461 gttatgcaga agaagactat gggatgggaa gcttcaagtg aaagaatgta ccctgaagac
     5521 ggtgctctta agggagagat taaacaacgt cttaaattga aagatggagg acattacgat
     5581 gctgaggtaa gtttaaacat gattttacta actaactaat ctgatttaaa ttttcaggtg
     5641 aagacaactt acaaagccaa aaaaccagtt cagctgccag gagcgtacaa tgttaatatt
     5701 aaactggata tcacctccca caacgaggat tacactatcg ttgagcaata tgaaagagct
     5761 gaagggcggc actcgacagg tggcatggat gaattgtata agtaggagct cccgagatct
     5821 tcagcgctca gcattcgtag aattgtaagt ttaaactttc tcatactaac taacgcgctc
     5881 tatttaaatt ttcagaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt
     5941 gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa
     6001 cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta
     6061 ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta
     6121 tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg
     6181 aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct
     6241 ccctcccccc attaataata attctatccc aaaatctaca caatgttctg tgtacacttc
     6301 ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa
     6361 taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg
     6421 ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt
     6481 tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc
     6541 ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg
     6601 agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta
     6661 gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat
     6721 acattcccaa tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt
     6781 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6841 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6901 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6961 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg
     7021 ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     7081 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7141 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7201 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7261 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7321 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7381 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7441 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7501 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7561 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7621 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7681 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7741 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7801 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7861 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7921 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7981 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8041 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8101 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8161 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8221 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     8281 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     8341 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     8401 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8461 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     8521 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     8581 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgag
//
