LOCUS       sequence-309326-        2591 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 2591)
  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 2591)
  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: pSEM323 - array co-injection fragment mosTI
            Pm...
FEATURES             Location/Qualifiers
     source          1..2591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(406..439)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(496..512)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(496..512)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(509..531)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(684..1272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(764..783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1389..1413)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1513..2304,2305..2373))
                     /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     2136..2155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2374..2478)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(2577..2591,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2591,1..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 cacgtgaata ggatgtgtct ggttttcagg ctttccctat ccaaaacgct
      121 cgttcgagtg tttcagttcg gtatgtcgag gcgttaccgg acggaaaata tttaaaataa
      181 aatcctcgtc atttcagatc ctaacgagaa gcgtgaccat atggttctct tagagtttgt
      241 aaccgccgct ggcataacac acggtatgga tgagctgtac aaatcacgaa gaagaaaggc
      301 taaccctact aaattgtcgg agaacgcaaa aaagttggcg aaagaagttg agaactagtc
      361 tgttcgttgt tccaataaat tgttattgtt ccctatttgg ctagcataac ttcgtatagc
      421 atacattata cgaagttata ggctaggtgg aggctcagtg atgataagtc tgcgatggtg
      481 gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcagagggca
      541 caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc agttcggcgt
      601 atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctct attcaacaaa
     1321 gccgccgtcc cgtcaagtca gcgtaaatgg gtagggggct tcaaatcgtc ctcgtgatac
     1381 caattcggag cctgcttttt tgtacaaact tgttgataat ggcaattcaa ggatcttcac
     1441 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1501 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1561 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1621 accatctggc cccagtgctg caatgatacc gcgagagcca cgctcaccgg ctccagattt
     1681 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1741 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1801 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1861 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1921 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1981 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2041 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2101 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2161 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     2221 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2281 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2341 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2401 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2461 acaaataggg gttccgcgca catttccccg aaaagtgcca gatacctgaa acaaaaccca
     2521 tcgtacggcc aaggaagtct ccaataactg tgatccacca caagcgccag ggttttccca
     2581 gtcacgacgt t
//
