LOCUS       sequence-361657-        4722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear mScarlet co-expression cloning vector for mosTI(unc-119). 
            Transgene inserted upstream of gpd-2 operon::mScarlet(2xNLS)::tbb-2 
            3'UTR. MCS or Golden Gate (BsaI: tgcc - MCS - tgag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4722)
  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 4722)
  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: pSEM399 - mosTI unc-119 - NNN - operon - mScar...
FEATURES             Location/Qualifiers
     source          1..4722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..34
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     26..43
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     27..43
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(667..700)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1029..1049
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1971..2294
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2653..2669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2653..2669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2666..2688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2841..3429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2921..2940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3546..3570)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3670..4461,4462..4530))
                     /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     4293..4312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4531..4635)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtcatgcat aatccgcacg
       61 catctggaat aaggaagtgc cattccgcct gacctaagtg cttttcgaga tcgccaaacc
      121 ggagaacgat caggagaatg atgagtcgcc acaggagtcg gcaagatacg tgcgttatag
      181 atttgctcca aactttttga aactcaaaac ggtcggagca actgtggaat tcaaagtagg
      241 agacatccca atccatcatt tccgaatgat cgaacgtcac ttcttcaaag atcgccttct
      301 gaagtgtttt gactttgaat tcggattctg tattccgaat tcacgaaaca actgtgaaca
      361 tatctatgag ttccctcaac tctctcaaca actcagtgag tcattattct aaaaagtaca
      421 attcaaaaga ctaatctctt tttcagtgga cgacatgatc aacaatccaa acgagactcg
      481 ttctgacagc ttctatttcg tcgataacaa actcgtcatg cacaacaaag ccgactactc
      541 atatgatgca taaatcttca ttgttgagtt tatcttgttg atttttgaat aaattatcaa
      601 ctctttactt tttaatgggt tatgaaataa ataaacattg aaaactgata aacaacgttc
      661 atctctataa cttcgtatag catacattat acgaagttat tgcctgagac cagctagcaa
      721 gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctca ggtctcttga
      781 gctgtctcat cctactttca cctagttaac tgcttgtctt aaaatctatg cttctcttta
      841 gtatctaaaa ttttcctaga agcttacaag tatataaatt gtctcttctc aataaaggtt
      901 gtatatttat tcatcttatt gaatctgcca tttcctcgtt tttgcgagtt tatatacctt
      961 ccaattttct ttctattgta ttttcaactt ctaattttaa ttcagggaaa ctgcttcaac
     1021 gcatcatgcc caaaaagaaa cgaaaggtcg tttctaaggg cgaagccgtc atcaaggtaa
     1081 gttttacgaa attctgaagc tcacaaaaca tggagtacat ttttcaggag ttcatgcggt
     1141 tcaaggtcca catggagggt agtatgaacg gacacgagtt cgagatagag ggagagggag
     1201 agggacggcc atacgaggga acccaaacgg ccaagctcaa ggtcaccaaa gggggaccac
     1261 tcccattctc ctgggatatc ctctccccac agttcatgta cggaagtcgt gccttcacca
     1321 agcaccccgc aggtaagttt ataatctatg atcttaattt ggcaatggaa aaagtttttt
     1381 cagatatacc agattactat aagcagtcct tcccagaggg attcaaatgg gagcgtgtca
     1441 tgaacttcga ggatggagga gccgtcaccg tcacgcaaga cacgtccctc gaggatggaa
     1501 ccctaatcta caaggtaaag cttcgtggaa ccaacttccc cccaggtaag ttttattcca
     1561 taatacaaat atatattgaa caataatgct tttcagatgg accagtcatg caaaagaaaa
     1621 ctatgggatg ggaagcctcg accgaacgtc tctacccaga ggacggggtc ctcaaaggag
     1681 atatcaagat ggccctaaga ctcaaggacg gaggacggta tcttgccgat ttcaagacca
     1741 cgtacaaggc taagaagcca gtacaaatgc caggggccta caacgtagac cgtaaactcg
     1801 acatcacgtc ccacaacgag gactacacgg tcgtcgaaca atacgaacgt tccgaaggac
     1861 gtcactccac cggagggatg gacgagctct acaagtcccg gcgtcgtaag gccaacccaa
     1921 cgaagctctc cgagaacgcc aaaaagctcg ccaaagaggt cgagaattaa atgcaaaatc
     1981 ctttcaagca ttcccttctt ctctatcact cttctttctt tttgtcaaaa aattctctcg
     2041 ctaatttatt tgctttttta atgttattat tttatgactt tttatagtca ctgaaaagtt
     2101 tgcatctgag tgaagtgaat gctatcaaaa tgtgattctg tctgatgtac tttcacaatc
     2161 tctcttcaat tccattttga agtgctttaa acccgaaagg ttgagaaaaa tgcgagcgct
     2221 caaatatttg tattgtgttc gttgagtgac ccaacaaaaa gaggaaactt tattgtgccg
     2281 ccaagaaaaa agtctcacag gaaacggaga atctgtgcgg caatgtctta tcatttgaaa
     2341 cgcgccgtat cgatgctgac atcatgcggc tcccgtgtta gtcgagaaga tgggtacggg
     2401 cctttattca tccaacactc tacggcttct tcaagagcta gtagggcacc ctgtagttgg
     2461 aaagggatta tttcgtatgg cgagcccata ccgtctttct tgcggaagct taacacgata
     2521 ggaagttgga tagtttcgaa cgatggttcc atacggaacg ctgtctcaag gatgagtctg
     2581 acggagtgta actcgaaggc taggtggagg ctcagtgatg ataagtctgc gatggtggat
     2641 gcatgtgtca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa
     2701 tcctattccg cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg
     2761 gcatatgtcg ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2821 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2881 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2941 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3001 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3061 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3241 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3361 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3421 ggatctcaag aagatccttt gatcttttct acggggtctg acgctctatt caacaaagcc
     3481 gccgtcccgt caagtcagcg taaatgggta gggggcttca aatcgtcctc gtgataccaa
     3541 ttcggagcct gcttttttgt acaaacttgt tgataatggc aattcaagga tcttcaccta
     3601 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3781 atctggcccc agtgctgcaa tgataccgcg agagccacgc tcaccggctc cagatttatc
     3841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3961 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4021 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4261 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4381 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4561 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4621 aataggggtt ccgcgcacat ttccccgaaa agtgccagat acctgaaaca aaacccatcg
     4681 tacggccaag gaagtctcca ataactgtga tccaccacaa gc
//
