LOCUS       sequence-174825-        7744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prgef-1-PH::miniSOG, expression of membrane targeted PH-miniSOG in 
            neurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7744)
  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 7744)
  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: pCZGY2839
FEATURES             Location/Qualifiers
     source          1..7744
                     /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(1651..1673)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1773..1792
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1870..1888)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1956..2060
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2061..2129,2130..2921)
                     /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(2279..2298)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3092..3680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3581..3600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3834..3851
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3968..3989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4004..4009,4010..4027,4028..4034)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4042..4058
                     /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     4047..4069
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4066..4082
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4066..4082
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    join(7737..7744,1..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 atgtaccatg gtattgatat ctgagctccg
      841 catcggccgc tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc taagtccaat
      901 tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat
      961 tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc ttttaagtca cctctaacaa
     1021 tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg
     1081 ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact
     1141 tcttatgttt tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa
     1201 aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc
     1261 tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa
     1321 tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc
     1381 ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga
     1441 tgagcacgat gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag
     1501 tagaagttga taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga
     1561 atacattccc aatataccaa acataactgt ttctactagt cggccgtacg ggccctttcg
     1621 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1681 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1741 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     1801 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     1861 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1921 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1981 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2041 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2101 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2161 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2221 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2281 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2341 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2401 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2461 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2521 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2581 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2641 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2701 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2761 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2821 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2881 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2941 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3001 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3061 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3121 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3181 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3241 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3301 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3361 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3421 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3481 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3541 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3601 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3661 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3721 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3781 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3841 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3901 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3961 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     4021 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     4081 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagcccccac acacatcgct
     4141 gccggaatag atgatcccat cgtagttcga taatctcgtg acactcgttt ccgatacccc
     4201 cttatatcag cacacattca gtcatcatac cacaaaatgt tgtgatagtg ataggcaaaa
     4261 ctaccgtatt tcctctatta accttgcatg caagactaat tttcgattaa cccgtagggg
     4321 tgcaagacta atagagactg caagactatt agaggctgaa atactaattt tcgtatgctc
     4381 aataattttg gaaattggcc tattttttgt agaaacttga taccgtttaa acaaggaaaa
     4441 atacacactt tttaatattt catcaataat ttgaacgatt ttgtgatttt aagttcaatt
     4501 tgcccaaaaa aagacaattt ttctgacgtt accgcaaagt aatgcaggtc aggcaaaaag
     4561 agcggtgcaa aaatataaga gactgcaata ctaatagagg aaatacggta attgaatttt
     4621 agtgaaactt gcgacagttt tcctcatttt tttgttattc cggcatacga gtgtggcata
     4681 cgagtgaggt catcttttgt ttcttccgtt tcttcatcgc ttttgaaaaa aaatgttgaa
     4741 gaaaacttcg tagccgatag ccggataatg tttggcacgg cgttccaaca tatatcattg
     4801 gaaatttttt aagatttttg cgaaaaatca cattcttcac gatgagaaca cgttattgaa
     4861 ggaatataaa tcaagaataa catatagtta tatttctcta ttactttaac gttaaatatg
     4921 agcaaatttg agcattttga ttgcgatgaa aagcagaatc gagtcaactg aaatccgttc
     4981 aaaaacctaa gctcggttcg cccgaaagtc gatttttcca atagccgaac agcactgctt
     5041 ctttattttc caggtatttt tggggtgtaa gaattatatc ttctttgttt gggtatttac
     5101 gtttccgaat tccctttctt gatctgtcgc tatttttcag aaaccagaac agtttatcct
     5161 ttttcatgag aacctgaata gctcaaaaac ccgtttaatt tcttttcatc atctcatttg
     5221 aaactttcct agtcttctta atgatttcct ggctcctcct acatttgtat ctcaagatgc
     5281 atacacttta tttccgctcc aattcgaatc atctacttct cttttttctt caactttcta
     5341 cttttttcca ttcttttctt tcaaatgttc gacgtcttca agtgactgct cttcattctc
     5401 ctcatttttc tcgaatttca ttcttgtctc ttttttgctc aaaaatggaa aatgaaagtg
     5461 acgtgagatt tggacggcgg gacacggggg cagtagaagc agcaaaaagg agagaaagag
     5521 gacacaaata agaagaacga attcaaaaat aagcggagag gagctatttc cgtcaattct
     5581 acctccccaa tcttcatcaa ttcggctcaa ttgaatgacg tcacagggag ataaaccgtt
     5641 tggatgagcg ccgtccatca ctgacgccat cccgtttggg acaagaaaag agaaaaaaga
     5701 gcacaaagtt tttggtgacg gatcttgtca atcatatgaa agttgttctg attgattgtc
     5761 agtttttttc ctactttttt ggattctatc cacttctgaa cttgtgacaa gtttcaaact
     5821 ttctgaatca ttttctatgc attttcctgg aattcttttt atgtaaaata tgaaatagaa
     5881 tgtttttgaa ttcaagttct gcttttttct tcttttttgt tctgttcggg cttgggtatg
     5941 ctttttttaa aaattatttt gcacatcgac caataagtgc gcaacttata aaattaattt
     6001 atttttgtta atttttgaaa tacttgtatt gctttaagtg atctgacctc gccctgagct
     6061 ttccacgtag ttatcaaata caaatcctcc aagggtaacg tacctatatt actgatcttt
     6121 ataataactt tatcacctgt ccagttccag aggaattctg ttaagcttat aggcacagaa
     6181 ggagtcattt ctgctggttt ttaatgatcc aaatctttat ttcaagtaaa aaactgaaca
     6241 cttgcgaata aaactatcag attaaccatt caccaaaaat gtgtttgaat ctaaaacttt
     6301 ctcagtattc caaatataga aataaataac cacgacattg ctaaaatctg tctgaattgt
     6361 gtactcctta ccgtgaagta ataatggata taatgaatcg tttgaaatga atgatcagca
     6421 catttttggt gaaagatcac aaataaggaa taagcgacgg aaaataaaac gattatttcg
     6481 gatcaaaaat ttgttgaaga tcatatacac tcgagaccaa gattattcta gacaattttc
     6541 aaattggctt ctttgtttgc aaatcttcat aataatctgt gaagtttgga aatttgaatt
     6601 tttaatcttt tttcatagat tatagttttt atttctttgc aaaactatat taaaaaccga
     6661 tgcattgttt tagggaaatt aatgagcctt ttgttcaaca ctaaaaacaa taaaactaga
     6721 attttggctt catcatttga ccttttttaa gttcgaaaac ttttctcgta tttctgaacc
     6781 gccaattttt tcacacatct ctagactttt ggtgcccgtt ccagaaagtt aagtaattgc
     6841 tattctaaga aagttctcaa cattgttttt tagttctgat tgaattctga tgttccagga
     6901 atatatttta aatttaatat tctttgcact atttctatac taactaaata ataaatagtc
     6961 taagtatgtt caatgagaac aaaaaagctc tatctatatt tttctatcct atttcattta
     7021 tatccttttc attttgaact caccagctcc tcttcttctt cttgtccata tacttcttaa
     7081 aagtctcgaa cttctctctc tcactctcca tcaatttctt tatggctacc gcttgcgctc
     7141 tgccgcttcc gaagaaagaa ggcgttggca gagctctcaa ttctattttt tctcgccgta
     7201 ggctatcgat ttctcagctt ctcttctctc ttcctcactc caccaccacc acttcgggct
     7261 tcttcttttc ttcattttcg tcttcttctt catcctcttc tttttttttc agttccccct
     7321 cactcctccc cttatatgcg cgtgcgagag gggtgcaaaa gcagcgcgca tccgagaatt
     7381 gaaacgagaa aacggagacg cagcagcagt tcgtcctcag aaaaatagcc agtaaaaaga
     7441 gaaaaagata gagagagacc tatcgcattt tattttcaaa ttgcaattcc tgcaattcat
     7501 gtgtgcgtgt gcccattttc aattcttccc gttatttttc agaccattac caacgttctt
     7561 tctagatctt gagacaatct tccttctgca gaatcaaggg cgaattccag cacactggcg
     7621 gccgttacta gaggatcccc gggattggcc aaaggaccca aaggtatgtt tcgaatgata
     7681 ctaacataac atagaacatt ttcaggagga cccttggcta gcgtcgacgg tacatcacaa
     7741 gttt
//
