LOCUS       sequence-490587-        6593 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  expression of GCaMP8f in C. elegans under myo2-promoter control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6593)
  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 6593)
  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: pLJ54 (Pmyo-2_GCaMP8f_SL2_mKate2_let-858_3'UTR)
FEATURES             Location/Qualifiers
     source          1..6593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1137..1154
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4163..4185)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4285..4304
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4386..4404)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4472..4576
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4577..4645,4646..5437)
                     /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(4795..4814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5608..6196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6097..6116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6350..6367
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6484..6505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6520..6525,6526..6543,6544..6550)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6558..6574
                     /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     6563..6585
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6582..6593,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6582..6593,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcg cggccgcact gactgggccg gcccatttta
       61 tatctgagta gtatcctttg ctttaaatgt ccataaaact aattttataa tcaataaaac
      121 aacgtttgta aatcaactga gtttacaagt agagacattg agggatactt tcactatgct
      181 aaagtgataa tcgaccaaat aataacttca ctttggtatt tattcctgtc ttataaatgt
      241 tatgtatgaa ttaaattcat atgcatatgg ctcactctga caatttttaa taatcttcca
      301 gatcaatatt gactaccgat gcgggtggtc ttttgctttg aattctgctg aactttacac
      361 cccgaacagc aatgtgtgct tcagcctaaa aaaaagtaag tgtgttaatc agtgcccccg
      421 attcttcatt ttttgcccct ctctcccgtt tcgtcggcaa aagaagagaa aataaagata
      481 agtctcaaga taggttggta atcgctaaag tggttgtgtg gataagagta gcaaaatggc
      541 aggaagagca ctttgcgcgc acacactgta ctcattgttc tggataaaat tctctcgttg
      601 tttgccgtcg gatgtctgcc tctctgcatt gagccggctt cttcactatc tttagttaac
      661 ctaaaatgcc gtttcttttc tcgtatcccc actatcccgt tgaggttctc tgctctcttc
      721 gctccctacc gccagcgagc aactatccgt gggggcgcct tgctcggaag atggggggga
      781 agaaagaaga tttttgctat ttgcacttga gaaagagact tttcctgcgt cgatggtcag
      841 agaacagtgt gcagacactt ttcagctacc tagaattaca attggatatc cccgcctccc
      901 aatccaccca cccagggaaa aagaagggct cgccgaaaat caaagttatc tccaggctcg
      961 cgcatcccac cgagcggttg acttctctcc accacttttc attttaaccc tcgatcgtca
     1021 gacacagaag gcgcgcctct agaggatccc cgggattggc caaaggaccc aaaggtatgt
     1081 ttcgaatgat actaacataa catagaacat tttcaggagg acccttggct agcatgcacc
     1141 accaccacca ccacactcgc cgtaagaaga ctttcaagga ggttgctacc gctgtcaaga
     1201 tcatcgctcg tcttatggga cttaagatta acgtttacat caaggctgat aagcaaaaga
     1261 acggaattaa ggctaacttc catatccgtc acaacatcga agatggagga gtccaacttg
     1321 cttaccatta ccaacaaaac actccaatcg gagatggacc agttcttctt ccagataacc
     1381 actacctttc cgttgagtcc aagctctcta aggtaagttt aaacatatat atactaacta
     1441 accctgatta tttaaatttt caggatccaa acgagaagcg tgaccacatg gtccttcttg
     1501 agttcgtcac cgctgctgga atcacccttg gaatggatga gctttataag ggaggaactg
     1561 gaggatctat ggtttctaag ggagaggagc ttttcactgg agttgtccca atccttgtcg
     1621 agcttgatgg agatgttaac ggacacaagt tctctgtttc tggagaggga gagggagacg
     1681 ctacctacgg aaagcttact cttaagttca tctgcactac tggaaagctt ccagttccat
     1741 ggccaaccct tgttaccacc cttacttacg gagttcaatg cttctctcgt tacccagatc
     1801 atatgaagca acatgatttc ttcaagtctg ctatgccaga gggatacatc caagagcgta
     1861 ctatcttctt caaggtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
     1921 ttcaggatga tggaaactac aagacccgtg ctgaggttaa gttcgaggga gatacccttg
     1981 ttaaccgtat cgagcttaag ggaatcgatt tcaaggaaga tggaaacatc cttggacata
     2041 agcttgagta caaccttcca gatcaactta ccgaggaaca aatcgctgag ttcaaggagg
     2101 ccttctccct tttcgataag gatggagacg gaactatcac taccaaggta agtttaaaca
     2161 tgattttact aactaactaa tctgatttaa attttcagga gcttggaacc gttatgcgtt
     2221 cccttggact taacccaacc gaggctgagc ttcaagatat gatcaacgag gttgacgctg
     2281 atggagatgg aactatcgat ttcccagagt tccttactat gatggctcgc aagatgaagt
     2341 accgcgatac tgaggaggag atccgcgagg ctttcggagt cttcgacaag gacggaaacg
     2401 gatacatttc cgctgctgag ctccgtcacg ttatgactaa cctcggagag aagcttaccg
     2461 acgaggaggt cgacgagatg atccgtgagg ctgatatcga cggagacgga caagttaact
     2521 acgaggagtt cgttcaaatg atgactgcca agtaaggtac cgctgtctca tcctactttc
     2581 acctagttaa ctgcttgtct taaaatctat gcttctcttt agtatctaaa attttcctag
     2641 aagcttacaa gtatataaat ggtctcttct caataaaggt tgtatattta ttcatcttat
     2701 tgaatctgcc atttcctcgt ttttgcgagt ttatatacct tccaattttc tttctattgt
     2761 attttcaact tctaatttta attcagggaa actgctgtac cggtagaaaa aatggtctcc
     2821 gagctcatta aagaaaacat gcatatgaag ctctacatgg agggaaccgt caacaaccac
     2881 cacttcaagt gcacctccga gggagaggga aagccatacg agggaaccca aaccatgcgt
     2941 atcaaggccg tcgagggagg accactccca ttcgccttcg acatcctcgc cacctccttc
     3001 atgtacggat ccaaggtaag tttaaacata tatatactaa ctaaccctga ttatttaaat
     3061 tttcagacct tcatcaacca cacccaagga atcccagact tcttcaagca atccttccca
     3121 gagggattca cctgggagcg tgtcaccacc tacgaggacg gaggagtcct caccgccacc
     3181 caagacacct ccctccaaga cggatgcctc atctacaacg tcaaggtaag tttaaacagt
     3241 tcggtactaa ctaaccatac atatttaaat tttcagatcc gtggagtcaa cttcccatcc
     3301 aacggaccag tcatgcaaaa gaagaccctc ggatgggagg cctccaccga gaccctctac
     3361 ccagccgacg gaggacttga gggacgtgcc gacatggccc tcaagctcgt cggaggagga
     3421 cacctcatct gcaacctcaa ggtaagttta aacatgattt tactaactaa ctaatctgat
     3481 ttaaattttc agaccaccta ccgttccaag aagccagcca agaacctcaa gatgccagga
     3541 gtctactacg tcgaccgtcg tcttgagcgt atcaaggagg ccgacaagga gacctacgtc
     3601 gagcaacacg aggtcgccgt cgcccgttac tgcgacctcc catccaagct cggacaccgt
     3661 taattcgcta gccggccata caagtaatcc ggatgatcga cgccaacgtc gttgaatttt
     3721 caaattttaa atactgaata tttgtttttt ttcctattat ttatttattc tctttgtgtt
     3781 ttttttcttg ctttctaaaa aattaattca atccaaatct aaacattttt ttttctcttt
     3841 ccgtctccca attcgtattc cgctcctctc atctgaacac aatgtgcaag tttatttatc
     3901 ttctcgcttt catttcatta ggacgtgggg ggaattggtg gaagggggaa acacacaaaa
     3961 ggatgatgga aatgaaataa ggacacacaa tatgcaacaa cattcaattc agaaatatgg
     4021 aggaaggttt aaaagaaaac ataaaaatat atagaggagg aaggaaaact agtaaaaaat
     4081 aagcaaagaa attaggcgaa cgatgagaat tgtcctcgct tgggcccttt cgtctcgcgc
     4141 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4201 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4261 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     4321 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cggccttaat
     4381 taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4441 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4501 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4561 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4621 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4681 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4741 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4801 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4861 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4921 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4981 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5041 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5101 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5161 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5221 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5281 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5341 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5401 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5461 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5521 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5581 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5641 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5701 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5761 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5821 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5881 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5941 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6001 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6061 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6301 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6361 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6421 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6481 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6541 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gct
//
