LOCUS       sequence-174819-        5245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pcol-19-PH::miniSOG(Q103L), expression of enhanced mutant version of
            membrane targeted PH-miniSOG (Q103L) in epidermis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5245)
  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 5245)
  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: pCZGY2850
FEATURES             Location/Qualifiers
     source          1..5245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     143..165
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     157..174
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     158..174
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     184..203
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        184..202
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1029..1053
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1812..1836)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1862..1878)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1862..1878)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2615..2635)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2615..2633)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2654..2670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2654..2670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2667..2689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2678..2694
                     /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(2702..2708,2709..2726,2727..2732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2747..2768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2885..2902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3056..3644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3136..3155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3815..4606,4607..4675))
                     /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     4438..4457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4676..4780)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(4806..5245,1..16))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4943..4964)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5155..5174
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
       61 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
      121 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
      181 gcgtaatacg actcactata gggcgaattg ggtaccagca tttgaaaatt tgcaccaatg
      241 tattatttaa tttttttttt cgaaaattta acgcattttc tctctaaaaa ctcgaaattt
      301 agtgtgttct aaacaacagt aagcatacaa tacctttgtt caaaattgac gtgctttctg
      361 aaccaatatg gttagtttca aaaatttttg tattatagga tagaaatatt tggaaataaa
      421 ttttaaaacc aaacttatgc ctttctcttt tagtatccca gctaggtaat attttagtat
      481 ttgcccaaat ccttgaagta aggagtatat aatttttgaa aaacaataaa actccagata
      541 attcatagtt ttttctcgaa agaaaatttt tgagattagt tattgaactt catttttgaa
      601 cattattcgt tgaaaaacac tcgctttgtc tttattttca aaaaaattcc gatttcccca
      661 accagaaaaa aaaacagata gaagaaattt ctccttaaat ttcattgtcc atctctcttg
      721 gaaacacatt atctatcaaa tgaaaaacgc attttttttc ctggcagaaa aatgaaattg
      781 gttagattac actggttagg tttgaaggtg taactttcgc tttctcagca actttcagta
      841 taaaaggaaa cggtcaccat ttagaaagac atcagttcat caactcgagg catgcctgca
      901 ggtcgactct agaggatccc cgggattggc caaaggaccc aaaggtatgt ttcgaatgat
      961 actaacataa catagaacat tttcaggagg acccttggag ggtacaggta gaaaaaaagc
     1021 ttgatatcac aagtttgtac aaaaaagcag gctccgaatt cgcccttcgg atccagatct
     1081 atgcacgggc tccaggatga cccggacctt caggcccttc tgaagggcag ccagcttctg
     1141 aaggtgaagt ccagctcgtg gcgtagggaa cgcttctaca agctacagga ggactgcaag
     1201 accatctggc aggaatctcg aaaggtcatg aggtccccgg agtcgcagct gttctccatc
     1261 gaggacattc aggaggtacg gatgggacac cgcacagaag gcctggagaa gtttgcccga
     1321 gacatccccg aggatcgatg cttctccatt gtcttcaagg accagcgcaa caccctagac
     1381 ctcattgccc catcaccagc tgacgctcag cactgggtgc agggcctgcg caagatcatc
     1441 caccactccg gctccatgga ccagcggcag aagactagtg agaaaagttt cgtgataact
     1501 gatccacggc tgccagacaa tcccatcatc ttcgcatccg atggcttcct ggagctgacc
     1561 gagtattcca gagaggagat cctgggccgc aatggccgct ttctgcaggg accagagaca
     1621 gaccaggcca cagtgcagaa gattcgcgat gccattagag atcagcgcga gattaccgtg
     1681 cagctgataa actacacaaa aagcgggaag aaattctgga acctcctgca cctccagccc
     1741 atgagggacc agaagggtga gctccagtat ttcatcggag tgctgctgga tggatgacaa
     1801 gggcgaattc gacccagctt tcttgtacaa agtggtgata tcgaattcct gcagcccggg
     1861 ggatccacta gttctagagc ggccgccacc gcggccgctg tcatcagatc gccatctcgc
     1921 gcccgtgcct ctgacttcta agtccaatta ctcttcaaca tccctacatg ctctttctcc
     1981 ctgtgctccc accccctatt tttgttatta tcaaaaaact tctcttaatt tctttgtttt
     2041 ttagcttctt ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa tagaattaat
     2101 tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata attctatccc
     2161 aaaatctaca caatgttctg tgtacacttc ttatgttttt tacttctgat aaattttttt
     2221 gaaacatcat agaaaaaacc gcacacaaaa taccttatca tatgttacgt ttcagtttat
     2281 gaccgcaatt tttatttctt cgcacgtctg ggcctctcat gacgtcaaat catgctcatc
     2341 gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa
     2401 ttttcctgct tttgcttttt ggggtttccc ctattgtttg tcaagatttc gaggacggcg
     2461 tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc ggaagaaggt
     2521 ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt ggagtagtgt
     2581 ctatggggtt tttgccgagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     2641 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2701 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2761 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2821 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2881 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2941 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3001 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3481 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3721 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4801 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4861 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4921 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4981 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5041 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5101 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5161 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5221 caccacaccc gccgcgctta atgcg
//
