LOCUS       sequence-174823-        5240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pmec-4-PH::miniSOG, expression of membrane targeted PH-miniSOG in 
            mechanosensory neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5240)
  AUTHORS   Xu S, Chisholm AD
  TITLE     Highly efficient optogenetic cell ablation in C. elegans using 
            membrane-targeted miniSOG.
  JOURNAL   Sci Rep. 2016 Feb 10;6:21271. doi: 10.1038/srep21271.
  PUBMED    26861262
REFERENCE   2  (bases 1 to 5240)
  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: "Sci Rep."; 
            date: "2016-02-10"; volume: "6:21271. doi"; pages: " 
            10.1038/srep21271"
COMMENT     Sequence Label: pCZGY2840
FEATURES             Location/Qualifiers
     source          1..5240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      152..740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     641..660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     894..911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1028..1049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1064..1069,1070..1087,1088..1094)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1102..1118
                     /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     1107..1129
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1126..1142
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1126..1142
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2293..2317
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2744..3061
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="EKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQGP
                     ETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQLD
                     G"
     protein_bind    complement(3085..3109)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3951..3973)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4073..4092
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4170..4188)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4256..4360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4361..4429,4430..5221)
                     /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(4579..4598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
       61 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      121 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      181 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      241 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      301 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      361 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      421 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      481 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      541 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      601 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      661 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      721 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      781 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
      841 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
      901 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
      961 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1021 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1081 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1141 accatgatta cgccaagctt caatacaagc tcaaatacaa tactattttt tttccgtctg
     1201 aacataaaat tagtttctaa ataaaaaatg tgaaaataaa tatttggttt taatcgttaa
     1261 aatcttccaa attactaata aagttcagtc aactatatgc ttcgaaattc actgaatcta
     1321 ttattgttat taatttttag acccttggag agctcccaac tctttcaatc cagaagactg
     1381 atggatgcca ggtgtacttg tcaaaggtgg ctcaaggatg cgagattgtc acttccaaat
     1441 ccagtgagat gaacatctcc gtgcaaacca atgatgatgg ggattatgta agtttttcgg
     1501 attgtagaac ccaatctaaa tttccttttc agtcggagtt cccggttccg gagcagttca
     1561 agacaacttt tgtcaatggc aagctagtca ctgtcgtctc ggacatttgc taaatcttct
     1621 cacgtcataa ccttattttt caacttcatg aactatcccc tcgaactttt ttatttgtcc
     1681 ctgtctacaa ttctttataa gcaattgaga cacactttgc tttattaaat aataccgatt
     1741 ttgttttgta tatgccttga ctggtgctca actattttta ttgaaaaaat tactttttac
     1801 tgtctcgtcg aataaaagtg atcccttcca aaaaatagtt attcatgttt gaaggtttgt
     1861 gatcgtagaa caattgcacc gaatgtagtg taaaaaaaat tccagaatgt tcaaatacat
     1921 ttccaccctc accagaacat ctcaaacacc aggcacattt caacacactt tcatggatct
     1981 tatcttgcct caaaaccatc agctttcttc tcattctaca agtatgaaac aatctactat
     2041 tctatacatt tataatgtaa tggaaccatg tgaagacatg agcaagatca ttgtcgtcag
     2101 cgactacccc tccttcattt ttcttagagt atcagtttgc taatgctatt ttttttatcg
     2161 ctatcaagtt atagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg
     2221 atactaacat aacatagaac attttcagga ggacccttgc ttggagggta ccgctagcgt
     2281 cgacggtaca tcacaagttt gtacaaaaaa gcaggctccg aattcgccct tcggatccag
     2341 atctatgcac gggctccagg atgacccgga ccttcaggcc cttctgaagg gcagccagct
     2401 tctgaaggtg aagtccagct cgtggcgtag ggaacgcttc tacaagctac aggaggactg
     2461 caagaccatc tggcaggaat ctcgaaaggt catgaggtcc ccggagtcgc agctgttctc
     2521 catcgaggac attcaggagg tacggatggg acaccgcaca gaaggcctgg agaagtttgc
     2581 ccgagacatc cccgaggatc gatgcttctc cattgtcttc aaggaccagc gcaacaccct
     2641 agacctcatt gccccatcac cagctgacgc tcagcactgg gtgcagggcc tgcgcaagat
     2701 catccaccac tccggctcca tggaccagcg gcagaagact agtgagaaaa gtttcgtgat
     2761 aactgatcca cggctgccag acaatcccat catcttcgca tccgatggct tcctggagct
     2821 gaccgagtat tccagagagg agatcctggg ccgcaatggc cgctttctgc agggaccaga
     2881 gacagaccag gccacagtgc agaagattcg cgatgccatt agagatcagc gcgagattac
     2941 cgtgcagctg ataaactaca caaaaagcgg gaagaaattc tggaacctcc tgcacctcca
     3001 gcccatgagg gaccagaagg gtgagctcca gtatttcatc ggagtgcagc tggatggatg
     3061 actcgagtaa caagggcgaa ttcgacccag ctttcttgta caaagtggtg atgtaccatg
     3121 gtattgatat ctgagctccg catcggccgc tgtcatcaga tcgccatctc gcgcccgtgc
     3181 ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     3241 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     3301 ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     3361 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     3421 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     3481 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     3541 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     3601 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     3661 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     3721 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     3781 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     3841 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttctactagt
     3901 cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3961 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4021 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4081 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     4141 agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa
     4201 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4261 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4321 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4381 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4441 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4501 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4561 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4621 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4681 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4741 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4801 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4861 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4921 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4981 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5041 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5101 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5161 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5221 actgtcagac caagtttact
//
