LOCUS       sequence-174821-        5254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pcol-19-PH::miniSOG, expression of membrane targeted PH-miniSOG in 
            epidermis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5254)
  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 5254)
  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: pCZGY2141
FEATURES             Location/Qualifiers
     source          1..5254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             444..761
                     /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(785..809)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(835..851)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(835..851)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1588..1608)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1588..1606)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1627..1643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1627..1643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1640..1662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1651..1667
                     /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)."
     promoter        complement(join(1675..1681,1682..1699,1700..1705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1720..1741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1858..1875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2029..2617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2109..2128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2788..3579,3580..3648))
                     /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     3411..3430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3649..3753)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3779..4234)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3916..3937)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4128..4147
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4361..4383
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4375..4392
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4376..4392
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4402..4421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4402..4420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(5247..5254,1..17)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gtacaaaaaa gcaggctccg aattcgccct tcggatccag atctatgcac gggctccagg
       61 atgacccgga ccttcaggcc cttctgaagg gcagccagct tctgaaggtg aagtccagct
      121 cgtggcgtag ggaacgcttc tacaagctac aggaggactg caagaccatc tggcaggaat
      181 ctcgaaaggt catgaggtcc ccggagtcgc agctgttctc catcgaggac attcaggagg
      241 tacggatggg acaccgcaca gaaggcctgg agaagtttgc ccgagacatc cccgaggatc
      301 gatgcttctc cattgtcttc aaggaccagc gcaacaccct agacctcatt gccccatcac
      361 cagctgacgc tcagcactgg gtgcagggcc tgcgcaagat catccaccac tccggctcca
      421 tggaccagcg gcagaagact agtgagaaaa gtttcgtgat aactgatcca cggctgccag
      481 acaatcccat catcttcgca tccgatggct tcctggagct gaccgagtat tccagagagg
      541 agatcctggg ccgcaatggc cgctttctgc agggaccaga gacagaccag gccacagtgc
      601 agaagattcg cgatgccatt agagatcagc gcgagattac cgtgcagctg ataaactaca
      661 caaaaagcgg gaagaaattc tggaacctcc tgcacctcca gcccatgagg gaccagaagg
      721 gtgagctcca gtatttcatc ggagtgcagc tggatggatg actcgagtaa caagggcgaa
      781 ttcgacccag ctttcttgta caaagtggtg atatcgaatt cctgcagccc gggggatcca
      841 ctagttctag agcggccgcc accgcggccg ctgtcatcag atcgccatct cgcgcccgtg
      901 cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc tccctgtgct
      961 cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt tttttagctt
     1021 cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt aattcgtaat
     1081 aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat cccaaaatct
     1141 acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt tttgaaacat
     1201 catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca
     1261 atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa
     1321 agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct
     1381 gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg gcgtttttct
     1441 tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt
     1501 tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg
     1561 gtttttgccg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     1621 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1681 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1741 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1801 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1861 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1921 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1981 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2041 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2101 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2161 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2221 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2281 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2341 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2401 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2461 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2521 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2581 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2641 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2701 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2761 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2821 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2881 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2941 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3001 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3061 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3121 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3181 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3241 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3301 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3361 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3421 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3481 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3541 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3601 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3661 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3721 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     3781 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3841 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3901 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3961 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4021 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4081 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4141 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4201 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     4261 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     4321 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     4381 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccagcatt
     4441 tgaaaatttg caccaatgta ttatttaatt ttttttttcg aaaatttaac gcattttctc
     4501 tctaaaaact cgaaatttag tgtgttctaa acaacagtaa gcatacaata cctttgttca
     4561 aaattgacgt gctttctgaa ccaatatggt tagtttcaaa aatttttgta ttataggata
     4621 gaaatatttg gaaataaatt ttaaaaccaa acttatgcct ttctctttta gtatcccagc
     4681 taggtaatat tttagtattt gcccaaatcc ttgaagtaag gagtatataa tttttgaaaa
     4741 acaataaaac tccagataat tcatagtttt ttctcgaaag aaaatttttg agattagtta
     4801 ttgaacttca tttttgaaca ttattcgttg aaaaacactc gctttgtctt tattttcaaa
     4861 aaaattccga tttccccaac cagaaaaaaa aacagataga agaaatttct ccttaaattt
     4921 cattgtccat ctctcttgga aacacattat ctatcaaatg aaaaacgcat tttttttcct
     4981 ggcagaaaaa tgaaattggt tagattacac tggttaggtt tgaaggtgta actttcgctt
     5041 tctcagcaac tttcagtata aaaggaaacg gtcaccattt agaaagacat cagttcatca
     5101 actcgaggca tgcctgcagg tcgactctag aggatccccg ggattggcca aaggacccaa
     5161 aggtatgttt cgaatgatac taacataaca tagaacattt tcaggaggac ccttggaggg
     5221 tacaggtaga aaaaaagctt gatatcacaa gttt
//
