LOCUS       sequence-317866-        2523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single copy or array insertion by MosTI using split Pmlc-2 GFP 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2523)
  AUTHORS   El Mouridi S, Alkhaldi F, Frokjaer-Jensen C
  TITLE     Modular safe-harbor transgene insertion (MosTI) for targeted 
            single-copy and extrachromosomal array integration in C. elegans.
  JOURNAL   G3 (Bethesda). 2022 Jul 28. pii: 6651068. doi: 
            10.1093/g3journal/jkac184.
  PUBMED    35900171
REFERENCE   2  (bases 1 to 2523)
  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: "G3 
            (Bethesda). 2022 Jul 28. pii: 6651068. doi: 
            10.1093/g3journal/jkac184."
COMMENT     Sequence Label: pSEM255 - sgRNA mosTI Pmlc-2_GFP
FEATURES             Location/Qualifiers
     source          1..2523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(428..444)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(428..444)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(441..463)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(616..1204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(696..715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1321..1345)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1445..2236,2237..2305))
                     /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     2068..2087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2306..2410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(2509..2523,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2523,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtcat gcataatccg cacgcatctg gaataaggaa gtgccattcc
       61 gcctgacctc gcctccccgc acactcaaca cagcgagcgg cggaacccgc ggggtgtcgg
      121 ccgctcggac acgcttcccg tatataacga cgcggcgcgc gtcaagttgt gcagacacat
      181 cctattcacg gtttaagagc tatgctggaa acagcatagc aagtttaaat aaggctagtc
      241 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tgttttttgc ccattttttg
      301 tagtttgaga ctggaaaaat cgtaaatcaa ccatagtttt agggcaaaat tctcgaaatt
      361 tttctaaaaa taggctaggt ggaggctcag tgatgataag tctgcgatgg tggatgcatg
      421 tgtcatggtc atagctgttt cctgtgtgaa attgttatcc gctcagaggg cacaatccta
      481 ttccgcgcta tccgacaatc tccaagacat taggtggagt tcagttcggc gtatggcata
      541 tgtcgctgga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      601 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      661 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      721 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      781 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      841 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      901 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      961 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1021 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1081 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1141 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1201 tcaagaagat cctttgatct tttctacggg gtctgacgct ctattcaaca aagccgccgt
     1261 cccgtcaagt cagcgtaaat gggtaggggg cttcaaatcg tcctcgtgat accaattcgg
     1321 agcctgcttt tttgtacaaa cttgttgata atggcaattc aaggatcttc acctagatcc
     1381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1561 gccccagtgc tgcaatgata ccgcgagagc cacgctcacc ggctccagat ttatcagcaa
     1621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2401 gggttccgcg cacatttccc cgaaaagtgc cagatacctg aaacaaaacc catcgtacgg
     2461 ccaaggaagt ctccaataac tgtgatccac cacaagcgcc agggttttcc cagtcacgac
     2521 gtt
//
