LOCUS       sequence-317416-        3207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single copy or array insertion by MosTI using split unc-119 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3207)
  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 3207)
  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: pSEM246 - MCS mosTI unc-119 target
FEATURES             Location/Qualifiers
     source          1..3207
                     /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(641..674)
                     /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(1112..1128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1112..1128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1125..1147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1300..1888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1380..1399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2005..2029)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2129..2920,2921..2989))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASQQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2752..2771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2990..3094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3193..3207,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3207,1..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 gcctgaccta agtgcttttc gagatcgcca aaccggagaa cgatcaggag aatgatgagt
      121 cgccacagga gtcggcaaga tacgtgcgtt atagatttgc tccaaacttt ttgaaactca
      181 aaacggtcgg agcaactgtg gaattcaaag taggagacat cccaatccat catttccgaa
      241 tgatcgaacg tcacttcttc aaagatcgcc ttctgaagtg ttttgacttt gaattcggat
      301 tctgtattcc gaattcacga aacaactgtg aacatatcta tgagttccct caactctctc
      361 aacaactcag tgagtcatta ttctaaaaag tacaattcaa aagactaatc tctttttcag
      421 tggacgacat gatcaacaat ccaaacgaga ctcgttctga cagcttctat ttcgtcgata
      481 acaaactcgt catgcacaac aaagccgact actcatatga tgcataaatc ttcattgttg
      541 agtttatctt gttgattttt gaataaatta tcaactcttt actttttaat gggttatgaa
      601 ataaataaac attgaaaact gataaacaac gttcatctct ataacttcgt atagcataca
      661 ttatacgaag ttattgcctg agaccagcta gcaagcttgc atgcctgcag gtcgactcta
      721 gaggatcccc gggtaccgag ctcaggtctc tgagctcagg aaacggagaa tctgtgcggc
      781 aatgtcttat catttgaaac gcgccgtatc gatgctgaca tcatgcggct cccgtgttag
      841 tcgagaagat gggtacgggc ctttattcat ccaacactct acggcttctt caagagctag
      901 tagggcaccc tgtagttgga aagggattat ttcgtatggc gagcccatac cgtctttctt
      961 gcggaagctt aacacgatag gaagttggat agtttcgaac gatggttcca tacggaacgc
     1021 tgtctcaagg atgagtctga cggagtgtaa ctcgaaggct aggtggaggc tcagtgatga
     1081 taagtctgcg atggtggatg catgtgtcat ggtcatagct gtttcctgtg tgaaattgtt
     1141 atccgctcag agggcacaat cctattccgc gctatccgac aatctccaag acattaggtg
     1201 gagttcagtt cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa aaggccagca
     1261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1741 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1921 cgctctattc aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag ggggcttcaa
     1981 atcgtcctcg tgataccaat tcggagcctg cttttttgta caaacttgtt gataatggca
     2041 attcaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2101 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2161 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2221 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gagccacgct
     2281 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2341 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgctgggaa gctagagtaa
     2401 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2461 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2521 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2581 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2641 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2701 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2761 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2821 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2881 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2941 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3001 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3061 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccagata
     3121 cctgaaacaa aacccatcgt acggccaagg aagtctccaa taactgtgat ccaccacaag
     3181 cgccagggtt ttcccagtca cgacgtt
//
