LOCUS       sequence-317437-        7929 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 7929)
  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 7929)
  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: pCFJ2560 - pEXP(mosTI, unc-119)[Peft-3 | ceGFP...
FEATURES             Location/Qualifiers
     source          1..7929
                     /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)."
     protein_bind    713..733
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        734..1333
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     protein_bind    1359..1383
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1392..1412
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4994..5065
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(5073..5097)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      5102..5425
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(5429..5449)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(5836..5852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5836..5852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5849..5871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(6024..6612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6104..6123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(6729..6753)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(6853..7644,7645..7713))
                     /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     7476..7495
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7714..7816)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(7915..7929,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7929,1..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 ttattgccat agcatggcct ttgcaggatt acgccaagct atcaactttg
      721 tatagaaaag ttggcacctt tggtctttta ttgtcaactt ccattggttc ttccattgtt
      781 tctgttaaat taatgaattt ttcataaaat aaagacatta tacaatataa aaatgaagaa
      841 tttattgaaa ataaactgcc agagagaaaa agtatgcaac actcccgccg agagtgtttg
      901 aaatggtgta cggtacattt tcgtgctagg agttagatgt gcaggcagca acgagagggg
      961 gagagatttt tttgggcctt gtgaaattaa cgtgagtttt ctggtcatct gactaatcat
     1021 gttggttttt tgttggttta ttttgttttt atctttgttt ttatccagat taggaaattt
     1081 aaattttatg aatttataat gaggtcaaac attcagtccc agcgtttttc ctgttctcac
     1141 tgtttagtcg aatttttatt ttaggctttc aacaaatgtt ctaactgtct tatttgtgac
     1201 ctcacttttt atattttttt aatttttaaa aatattagaa gtttctagga taattttttc
     1261 gacttttatt ctctctaccg tccgcactct tcttactttt aaattaaatt gttttttttt
     1321 cagttgggaa acactttgct cactccgtag cagccatggc aagtttgtac aaaaaagcag
     1381 gctcgggatc cccaaagaag aagcgtaagg tgtccaaggt aacacttagt ttttgttgct
     1441 ctgtttaaaa attaatttta agggagagga gctgttcacc ggagtcgtcc caatcctcgt
     1501 cgagctcgac ggggacgtca acggacacaa gttctccgtg tccggagagg gagagggaga
     1561 tgccacctac ggaaagctca ccctcaagtt catctgcacc accggaaagc tcccagtccc
     1621 atggccaacc ctcgtcacca ccttctgcta cggagtccaa tgcttctccc gttacccagg
     1681 taagttttct gttttattta cttgttttga gaattgggat ttttagctga aaaattggat
     1741 tttaagatga aaaattgaaa ttttcaacta aaaatttcaa tttttagctg aaaagttaaa
     1801 tttttagcta aagagtggga tttttagttg aaaaatcgag atttctagct aaaaagtatg
     1861 atttttagct gaaagattgg aattttaagc agaaaaaatt gtatttttag ctgataaatc
     1921 aaattttaag cggaaaaatt tgaattttta gctaaaaaat tccattttca actaaaaatc
     1981 gtgattttta gctttaaaaa ttggattttt agctgaaaaa tcggattttt agctgaaaat
     2041 tgaaattttt tagttaaaaa gtgagatttt ttaacagaaa agtgagattt ttagttaaaa
     2101 aataatattt ttagctaaaa aatacaattt tcaactaaag atcgtgattt ttagctttaa
     2161 aaattggatt tttagctaaa aaatcggatt tttagctgaa aattgaaatt ttttagttaa
     2221 aaagtgggat tttaagatga aaagtgagat tttttagctg aaaagtgaga ttttcagcta
     2281 aaaaatacaa ttttcaactg aaaaattggg atttttagct ttaaaaattg gattttcagc
     2341 tttaaaaatt ggatttttag ctgaaaagtg agatttttag ttgaaaaatt gaaatgttta
     2401 gctgaaaaat tggaattttt aagtaaaaaa gtgggatttt aagatgaaaa gtgagatgtt
     2461 tttagctgaa aagtgagatt tttagctaaa aaatacaatt ttcaactgaa aaattgggat
     2521 ttttagctca aaagtgggac gaaagacgaa aaaaaaaatc gattttttac caaaaattag
     2581 attttcagac cacatgaagc gtcacgactt cttcaagtcc gccatgccag agggatacgt
     2641 ccaagagcgt accatcttct tcaaggacga cggaaactac aaggtaagtt atgtacgcga
     2701 tttctcggcg aaaaaatgga cttttttgct caaaaactga atttttcagc tttttttgca
     2761 ctgaaaatgc tgaaaattgg tttttttgct gaaaattcac cgaaaaaagc tgaaaatatt
     2821 gcagaaatcg acgaaaaatg ccttttttgc tgaaatttca gccagaaaat tgttcagaat
     2881 cgcaatttcc tgccaaaatc gacgtttttc ggcgaaaatt ggtagatttt gaaccgaaaa
     2941 ttgtgaaaaa tggacttttt tgctcaaaaa cagacttttt gagctttttt cacactgaaa
     3001 atggtggaaa attggatttt ccagcagaaa attcggaaat tttcaatttt tggcggaaat
     3061 ttgacgaaaa atatatgaaa aacaccgatt tttgtctgaa aaagttcaaa agccgaaaaa
     3121 ttggccaaaa atctccaaaa tcggactttt ctggcgtttt tcgtggaaaa attgaaattt
     3181 ttttgaaaaa ttgagcaaaa atcgccgaaa aaccgctaaa aaaactgccg aaattggcaa
     3241 ttttgagctt aaaaaaactt gaaaaatgct caaaattacc caattttcac cggaaattga
     3301 gctaaaatcg tcgaaaaaga ctgaattttc aatttttagc tgaaaatctg cctattttcg
     3361 gctaaaattg ctttaaaaac attaaaattc aaattttttg gctaaaaata ctcagattgc
     3421 cctttttttt ctccaaaaat gacaaaaaaa aaatcaatca gaaaaatgcg atttttttta
     3481 caaaaattgg cttcaaaacg tctaaattct ccctaatttt tgctgctgaa aattgtgcaa
     3541 aaatcaccga aaaatgacga ttttaggctg aaaatcttca aaaaaagctc aaaattcccc
     3601 acttttcacc ggaaaaagac gaaaaaagct ccaaatttat cgattaacat tttcagaccc
     3661 gtgccgaggt caagttcgag ggagacaccc tcgtcaaccg tatcgagctc aagggaatcg
     3721 acttcaagga ggacggtaac atcctgggac acaagctgga gtacaactac aactcccaca
     3781 acgtctacat catggccgac aagcaaaaga acggaatcaa ggtcaacttc aaggtaagtt
     3841 cttttttttg ctcgtttttg agtgaaaaat tgatggaatt cacaaaaaat ggccaaagaa
     3901 tcctcaaaat tggccgaaaa ttctgaaaaa tgacctaaaa atcaccgaaa aacggccaaa
     3961 aattatctga aaatcgccga aaatctcaaa ttttgaggaa attcccgcta aaaattaata
     4021 aaaatcaccg aaaaaaggcc aaaaattacc taataaaggc cgaaaatcgc ctaaaaaggc
     4081 caaaaattct gaaaaatggc caaatttcaa tttttcacac acaaaaaatc cctgaatttt
     4141 ctatttttac tcgaaaaact gtggatttac gggggtaaaa attcagtttt tagtgctatt
     4201 tcacaagaaa aacacagcat tttcgacaaa aactccgatt tttaagccaa aaatttgagg
     4261 tttttagttt tttacgtgaa aacctgaact ttttgccaaa aaaacacttt tttttagact
     4321 aaaaatcaat tttcgatcaa aaatcactaa ttttgcctga aaaaatatag atttttccac
     4381 tttttcactg aaaaaaaact ggattttcac tagaaaatca agttgttttt agaccaaaaa
     4441 ttttattttt ccgctgaaaa ttatcgtttt tcagtcaatt ttcagctaaa aatccctgaa
     4501 attctggatt ttcagccaaa aaactcgaaa ttttccaaaa aaaccccgtt taaaaatgag
     4561 tttagactga aaattccgat ttttgagcca aaaatttgat ttttaatcga aaaaaattgg
     4621 attttcaacc aaaaaatcag tttttcagtg atttttagag taaaaatcac tgattttaac
     4681 acaaaattcc agtttttcgt gatttttcac tagaaaatcc cgatttttcc ccaaaaaatc
     4741 catttttccg actaaaaatc cccaaagtaa ttttcagatc cgtcacaaca tcgaggacgg
     4801 atcggtccaa ctcgccgacc actaccaaca aaacacccca atcggtgacg gaccagtcct
     4861 cctcccagac aaccactacc tctccaccca atccgccctc tccaaggacc caaacgagaa
     4921 gcgtgaccac atggtcctcc tcgagttcgt caccgccgcc ggaatcaccc acggaatgga
     4981 cgagctctac aagtcccgtc gtcgtaaggc caacccaacc aagctctccg agaacgccaa
     5041 gaagctcgcc aaggaggtcg agaacggatc caacccagct ttcttgtaca aagtgggata
     5101 aatgcaaaat cctttcaagc attcccttct tctctatcac tcttctttct ttttgtcaaa
     5161 aaattctctc gctaatttat ttgctttttt aatgttatta ttttatgact ttttatagtc
     5221 actgaaaagt ttgcatctga gtgaagtgaa tgctatcaaa atgtgattct gtctgatgta
     5281 ctttcacaat ctctcttcaa ttccattttg aagtgcttta aacccgaaag gttgagaaaa
     5341 atgcgagcgc tcaaatattt gtattgtgtt cgttgagtga cccaacaaaa agaggaaact
     5401 ttattgtgcc gccaagaaaa aagtctcaca actttattat acatagttga taattccctc
     5461 tgacacatgc agctcgagct caggaaacgg agaatctgtg cggcaatgtc ttatcatttg
     5521 aaacgcgccg tatcgatgct gacatcatgc ggctcccgtg ttagtcgaga agatgggtac
     5581 gggcctttat tcatccaaca ctctacggct tcttcaagag ctagtagggc accctgtagt
     5641 tggaaaggga ttatttcgta tggcgagccc ataccgtctt tcttgcggaa gcttaacacg
     5701 ataggaagtt ggatagtttc gaacgatggt tccatacgga acgctgtctc aaggatgagt
     5761 ctgacggagt gtaactcgaa ggctaggtgg aggctcagtg atgataagtc tgcgatggtg
     5821 gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcagagggca
     5881 caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc agttcggcgt
     5941 atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6001 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6061 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6421 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctct attcaacaaa
     6661 gccgccgtcc cgtcaagtca gcgtaaatgg gtagggggct tcaaatcgtc ctcgtgatac
     6721 caattcggag cctgcttttt tgtacaaact tgttgataat ggcaattcaa ggatcttcac
     6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6961 accatctggc cccagtgctg caatgatacc gcgagagcca cgctcaccgg ctccagattt
     7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7081 cgcctccatc cagtctatta attgttgctg ggaagctaga gtaagtagtt cgccagttaa
     7141 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7441 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7741 catttatcag ggttattgtc tcatgagcgg atacatttga atgtatttag aaaaataaac
     7801 aaataggggt tccgcgcaca tttccccgaa aagtgccaga tacctgaaac aaaacccatc
     7861 gtacggccaa ggaagtctcc aataactgtg atccaccaca agcgccaggg ttttcccagt
     7921 cacgacgtt
//
