LOCUS       sequence-174822-        5665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pnmr-1-PH::miniSOG, expression of membrane targeted PH-miniSOG in 
            subset of interneurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5665)
  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 5665)
  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: pCZGY2845
FEATURES             Location/Qualifiers
     source          1..5665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1882..1906
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2333..2650
                     /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(2674..2698)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2724..2740)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2724..2740)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3477..3497)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3477..3495)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3516..3532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3516..3532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3529..3551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3540..3556
                     /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(3564..3570,3571..3588,3589..3594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3609..3630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3747..3764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3918..4506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3998..4017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4677..5468,5469..5537))
                     /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     5300..5319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5538..5642)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacctt
      661 ctgaaagtat aagatttgac tgaaaattat aaaattatga acttgagaaa gtctagaaat
      721 caagaaaaaa tatctagaaa tgaacattcc atgggttgtg aagatgcttt tatctcactt
      781 tagaatgtct gactccgccc ctctgtgtac gaattgaagt gctcttttgt gatgatcata
      841 gctcctcctt gcggcgacgc ccgcacacag cacgagatat tacacggaat acttatgtga
      901 ttcataggaa acaaggaaat cgaaaaaata ataggatttt gaggaggtat ttttagatga
      961 atgcttatga tgttatgatg aacccaagaa attctgaata gacatatgtc agaacggggt
     1021 aggagctcca gtgtcgcttt tgaaataaaa aatcaagtgg tatacttctc gtatataccc
     1081 ttctgaacta ataaaataat ttatcttttg aaaaagtttg aaaaagaaaa atgcccaaaa
     1141 cttgaatttt aatgttttgt attttgagca tatcgttcaa acgtcgtgat ctgaaagtag
     1201 aaagatctaa atattcgtga cacagtttgg tttgaaggtc ttagtggtaa ttttttatta
     1261 cagaagcaac aaaaagttga catattttca gaatttccaa aaaaacgttt aaaatttcaa
     1321 tttcttcgat agagttctaa aaattttttg gaacggattt tgtaatatcc gctgataacc
     1381 gttcaagaac aatttgaatt ttgaaataat ttaatcaact agtatccagt gctctttctc
     1441 actgctccac ccatttttca acccatttcc tttactcaat ttcgcccgtt ttccttttgc
     1501 tcaaatgtgc ccaccaaatc atctttcttc gtcttctttt ctcttattcg gaaggagaag
     1561 aggtgaaagt agaatacgtt tttttgttga aaagagaaga gcactgccac ttttctctta
     1621 tttccatcct tctactacct cgttttactt tccatttaaa aacccgttta aagttccaaa
     1681 atttcaatta tcttgcttct gtttggaaca cgacaaactt tagttttgtt acagataagc
     1741 ttgcatgcct gcaggtcgac tctagaggat ccccgggatt ggccaaagga cccaaaggta
     1801 tgtttcgaat gatactaaca taacatagaa cattttcagg aggacccttg gagggtaccg
     1861 gtagaaaaaa agcttgatat cacaagtttg tacaaaaaag caggctccga attcgccctt
     1921 cggatccaga tctatgcacg ggctccagga tgacccggac cttcaggccc ttctgaaggg
     1981 cagccagctt ctgaaggtga agtccagctc gtggcgtagg gaacgcttct acaagctaca
     2041 ggaggactgc aagaccatct ggcaggaatc tcgaaaggtc atgaggtccc cggagtcgca
     2101 gctgttctcc atcgaggaca ttcaggaggt acggatggga caccgcacag aaggcctgga
     2161 gaagtttgcc cgagacatcc ccgaggatcg atgcttctcc attgtcttca aggaccagcg
     2221 caacacccta gacctcattg ccccatcacc agctgacgct cagcactggg tgcagggcct
     2281 gcgcaagatc atccaccact ccggctccat ggaccagcgg cagaagacta gtgagaaaag
     2341 tttcgtgata actgatccac ggctgccaga caatcccatc atcttcgcat ccgatggctt
     2401 cctggagctg accgagtatt ccagagagga gatcctgggc cgcaatggcc gctttctgca
     2461 gggaccagag acagaccagg ccacagtgca gaagattcgc gatgccatta gagatcagcg
     2521 cgagattacc gtgcagctga taaactacac aaaaagcggg aagaaattct ggaacctcct
     2581 gcacctccag cccatgaggg accagaaggg tgagctccag tatttcatcg gagtgcagct
     2641 ggatggatga ctcgagtaac aagggcgaat tcgacccagc tttcttgtac aaagtggtga
     2701 tatcgaattc ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggccgc
     2761 tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc taagtccaat tactcttcaa
     2821 catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat tatcaaaaaa
     2881 cttctcttaa tttctttgtt ttttagcttc ttttaagtca cctctaacaa tgaaattgtg
     2941 tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc
     3001 ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact tcttatgttt
     3061 tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa aataccttat
     3121 catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc
     3181 atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa tttttcaatc
     3241 aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc ccctattgtt
     3301 tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat
     3361 gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga
     3421 taatttgaaa gtggagtagt gtctatgggg tttttgccga gctccagctt ttgttccctt
     3481 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3541 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3601 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3661 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3721 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3781 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3841 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3901 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3961 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4021 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4081 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4141 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4201 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4261 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4321 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4381 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4441 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4501 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4561 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4621 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4681 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4741 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4801 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4861 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4921 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4981 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5041 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5101 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5161 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5221 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5281 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5341 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5401 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5461 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5521 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5581 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5641 cgcacatttc cccgaaaagt gccac
//
