LOCUS       sequence-490588-        7588 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  expression of GCaMP8f in C. elegans under mec-17-promoter control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7588)
  AUTHORS   Liu J, Bonnard E, Scholz M
  TITLE     Adapting and optimizing GCaMP8f for use in Caenorhabditis elegans.
  JOURNAL   Genetics. 2024 Oct 7;228(2):iyae125. doi: 10.1093/genetics/iyae125.
  PUBMED    39074213
REFERENCE   2  (bases 1 to 7588)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics.";
            date: "2024-10-7"; volume: "228(2):iyae125. doi"; pages: " 
            10.1093/genetics/iyae125"
COMMENT     Sequence Label: pLJ57 (Pmec-17_GCaMP8f_SL2_mKate2_let-858_3'UTR)
FEATURES             Location/Qualifiers
     source          1..7588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2132..2149
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5158..5180)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5280..5299
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5381..5399)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5467..5571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5572..5640,5641..6432)
                     /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(5790..5809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6603..7191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7092..7111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7345..7362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7479..7500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7515..7520,7521..7538,7539..7545)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7553..7569
                     /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     7558..7580
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7577..7588,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7577..7588,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcg cggccgcact gactgggccg gccactactt
       61 gagaggtccg gcaacttgtt tgaccacttt tgcaaatttt tgatgctttt ctggatataa
      121 ttaggtgacc attcttcagg tagggtaaac attctggaag ctaggtttta taagatgaaa
      181 acaaaacaca cttgccacga agcttccaga atgtttgtaa aaatctaatt tggaagagtt
      241 atatttgtga gcggaaataa tttccttata attttcagct tgtaaaatca ttcacatcta
      301 ataaaaagca aattaggtaa actataaaat taacaaataa attaccactg cttgaagtgt
      361 agatgtgata agaattggta gagcaaagac ggcaagaact gcaacaaata cagcaactga
      421 tgacgagaaa acggccaatt gacgcattcg tcgcgctaac agttctagat ctttctcatt
      481 acccttcatt ttctgaaatt actattataa atgtgttagc caagagtctc ccattgatca
      541 cagaatggca aactcaaatt ataaaatata tctgaaacag attaaatgtt attgtataag
      601 atttacattt ttcaagtttt aatttccatt attgtttccg tttttctgtc aagaaatcta
      661 tagatgtgag catcaaattt ttacaataag tttgttggag agacttttaa aaatgaatac
      721 aataattgaa tttttcttca aagtgaggaa aaaagtaaat gttgtatccg aatgaaagtt
      781 caatacattg aaaaataatg tgaatgaaaa gcattcaaat gttcagattt tcgttcaata
      841 cgaggctaac agcataaaaa aaagaatacg cgttatcaca gtgttttaag tgtaaaagag
      901 tgacgaagtc taatgaaaag gtatgcgtag tttacaatgg ataatttggg tgtggagaag
      961 ttagaatgat gtctgctgat gttaatcact tgaattcttg ttttctgatc tgcaaaagct
     1021 tcttcttgaa atggttctcg ttcttgatat tcgagatagc tttgaaaatt ccaaattttt
     1081 gacaaacatc tagaaattta tttgaagatg aagcagtgtc tgtagaaata gtaatcatgt
     1141 tttcaatatt tatcatgttt tcaataccgt ccaacatggc caaatattat aaaaaactct
     1201 gaagtgtaaa aaaatatttt tctatttttg aaaattttgg caaattccca attttcgaga
     1261 aatattggtt tttaaaaaat aattttagag caatgtcgca tttttgagca tctacaatgt
     1321 tcagatgcaa ataattaaaa tataaacttc caaaattaaa agtgagagga actgagtggt
     1381 tgacactttt ggctgtaaaa ttagcactat gtagaagagt ggtgttgaaa aacacaaagt
     1441 gtcttgtaaa gaaatctaca gtttcagaaa acctggatat tcaaagtttc tgaaaagtac
     1501 cactcaaatt gctcctagaa ataattttaa aaaacgattc tccattttct gataaaattt
     1561 caagttacgg aatgtttcaa attctaaatg aatctgatta tttttcttgc tacacgaatc
     1621 tgaaaatcta aaaatagtct ttttcaaagt tccaatgaaa tttccagaga tctcgaattt
     1681 agattttttg gaacagtaat taaaaatgtt tatctgtttg tgactcttcc aacttcttcc
     1741 gcgtttgtgt tttcctttct aaattctgcc aaaaaagaaa aagatcatcc gaatgtttca
     1801 gaaatgaaga gtcgtcaaga aaagaaataa tgccaaacac tgatataaaa ttcatagaaa
     1861 agtcactttg tatactgtat gaattccgaa gaaattgaaa ggaattctcc gataagagca
     1921 ggctcaccaa aaatgcatga gaatggggag gaaatgtggg aggaggagag ctgagatgaa
     1981 gagaagctgc ttcaccgatg gatcagttgt gagacgattc gatcggcgcg cctctagagg
     2041 atccccggga ttggccaaag gacccaaagg tatgtttcga atgatactaa cataacatag
     2101 aacattttca ggaggaccct tggctagcat gcaccaccac caccaccaca ctcgccgtaa
     2161 gaagactttc aaggaggttg ctaccgctgt caagatcatc gctcgtctta tgggacttaa
     2221 gattaacgtt tacatcaagg ctgataagca aaagaacgga attaaggcta acttccatat
     2281 ccgtcacaac atcgaagatg gaggagtcca acttgcttac cattaccaac aaaacactcc
     2341 aatcggagat ggaccagttc ttcttccaga taaccactac ctttccgttg agtccaagct
     2401 ctctaaggta agtttaaaca tatatatact aactaaccct gattatttaa attttcagga
     2461 tccaaacgag aagcgtgacc acatggtcct tcttgagttc gtcaccgctg ctggaatcac
     2521 ccttggaatg gatgagcttt ataagggagg aactggagga tctatggttt ctaagggaga
     2581 ggagcttttc actggagttg tcccaatcct tgtcgagctt gatggagatg ttaacggaca
     2641 caagttctct gtttctggag agggagaggg agacgctacc tacggaaagc ttactcttaa
     2701 gttcatctgc actactggaa agcttccagt tccatggcca acccttgtta ccacccttac
     2761 ttacggagtt caatgcttct ctcgttaccc agatcatatg aagcaacatg atttcttcaa
     2821 gtctgctatg ccagagggat acatccaaga gcgtactatc ttcttcaagg taagtttaaa
     2881 cagttcggta ctaactaacc atacatattt aaattttcag gatgatggaa actacaagac
     2941 ccgtgctgag gttaagttcg agggagatac ccttgttaac cgtatcgagc ttaagggaat
     3001 cgatttcaag gaagatggaa acatccttgg acataagctt gagtacaacc ttccagatca
     3061 acttaccgag gaacaaatcg ctgagttcaa ggaggccttc tcccttttcg ataaggatgg
     3121 agacggaact atcactacca aggtaagttt aaacatgatt ttactaacta actaatctga
     3181 tttaaatttt caggagcttg gaaccgttat gcgttccctt ggacttaacc caaccgaggc
     3241 tgagcttcaa gatatgatca acgaggttga cgctgatgga gatggaacta tcgatttccc
     3301 agagttcctt actatgatgg ctcgcaagat gaagtaccgc gatactgagg aggagatccg
     3361 cgaggctttc ggagtcttcg acaaggacgg aaacggatac atttccgctg ctgagctccg
     3421 tcacgttatg actaacctcg gagagaagct taccgacgag gaggtcgacg agatgatccg
     3481 tgaggctgat atcgacggag acggacaagt taactacgag gagttcgttc aaatgatgac
     3541 tgccaagtaa ggtaccgctg tctcatccta ctttcaccta gttaactgct tgtcttaaaa
     3601 tctatgcttc tctttagtat ctaaaatttt cctagaagct tacaagtata taaatggtct
     3661 cttctcaata aaggttgtat atttattcat cttattgaat ctgccatttc ctcgtttttg
     3721 cgagtttata taccttccaa ttttctttct attgtatttt caacttctaa ttttaattca
     3781 gggaaactgc tgtaccggta gaaaaaatgg tctccgagct cattaaagaa aacatgcata
     3841 tgaagctcta catggaggga accgtcaaca accaccactt caagtgcacc tccgagggag
     3901 agggaaagcc atacgaggga acccaaacca tgcgtatcaa ggccgtcgag ggaggaccac
     3961 tcccattcgc cttcgacatc ctcgccacct ccttcatgta cggatccaag gtaagtttaa
     4021 acatatatat actaactaac cctgattatt taaattttca gaccttcatc aaccacaccc
     4081 aaggaatccc agacttcttc aagcaatcct tcccagaggg attcacctgg gagcgtgtca
     4141 ccacctacga ggacggagga gtcctcaccg ccacccaaga cacctccctc caagacggat
     4201 gcctcatcta caacgtcaag gtaagtttaa acagttcggt actaactaac catacatatt
     4261 taaattttca gatccgtgga gtcaacttcc catccaacgg accagtcatg caaaagaaga
     4321 ccctcggatg ggaggcctcc accgagaccc tctacccagc cgacggagga cttgagggac
     4381 gtgccgacat ggccctcaag ctcgtcggag gaggacacct catctgcaac ctcaaggtaa
     4441 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagacc acctaccgtt
     4501 ccaagaagcc agccaagaac ctcaagatgc caggagtcta ctacgtcgac cgtcgtcttg
     4561 agcgtatcaa ggaggccgac aaggagacct acgtcgagca acacgaggtc gccgtcgccc
     4621 gttactgcga cctcccatcc aagctcggac accgttaatt cgctagccgg ccatacaagt
     4681 aatccggatg atcgacgcca acgtcgttga attttcaaat tttaaatact gaatatttgt
     4741 tttttttcct attatttatt tattctcttt gtgttttttt tcttgctttc taaaaaatta
     4801 attcaatcca aatctaaaca tttttttttc tctttccgtc tcccaattcg tattccgctc
     4861 ctctcatctg aacacaatgt gcaagtttat ttatcttctc gctttcattt cattaggacg
     4921 tggggggaat tggtggaagg gggaaacaca caaaaggatg atggaaatga aataaggaca
     4981 cacaatatgc aacaacattc aattcagaaa tatggaggaa ggtttaaaag aaaacataaa
     5041 aatatataga ggaggaagga aaactagtaa aaaataagca aagaaattag gcgaacgatg
     5101 agaattgtcc tcgcttgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5161 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5221 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5281 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5341 cgtaaggaga aaataccgca tcaggcggcc ttaattaagg gcctcgtgat acgcctattt
     5401 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6721 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7261 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7321 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7381 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7441 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7501 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7561 ggataacaat ttcacacagg aaacagct
//
