LOCUS       sequence-361680-        4403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP fusion cloning vector for mosTI(unc-119). Transgene inserted 
            upstream of gfp and tbb-2 3' UTR. MCS or Golden Gate (BsaI: tgcc - 
            MCS - aaaa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4403)
  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 4403)
  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: pSEM386 - mosTI unc-119 - NNN - gfp - tbb-2
FEATURES             Location/Qualifiers
     source          1..4403
                     /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)."
     terminator      1626..1949
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2308..2324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2308..2324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2321..2343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2496..3084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2576..2595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3201..3225)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3325..4116,4117..4185))
                     /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     3948..3967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4186..4290)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(4389..4403,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4403,1..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 taaaaatgtc aaaaggagaa gaactcttta
      781 ccggagtcgt cccaattctc gtcgagctcg acggagacgt aaacggacac aaattctcgg
      841 tttccggaga aggagaagga ggtaagtttt acgaaattct gaagctcaca aaacatggag
      901 tacatttttc agatgctact tatggaaaac tcacccttaa attcatttgc accaccggaa
      961 agctcccagt gccgtggccg acccttgtga ctacattctg ctacggagtt caatgctttt
     1021 cccgctaccc agaccacatg aagcgtcacg atttcttcaa atcagccatg ccagaaggat
     1081 acgtccagga gcgaacaatt ttcttcaagg acgatggaaa ctacaagact cgtgctgaag
     1141 tcaagtttga aggagatact ctcgtgaacc gcattgagct caaggtaagt ttataatcta
     1201 tgatcttaat ttggcaatgg aaaaagtttt ttcagggaat cgacttcaaa gaagatggaa
     1261 acattcttgg acacaagctc gaatacaact ataactcgca caacgtgtat atcatggccg
     1321 acaagcaaaa aaatggaatc aaggtcaact tcaagattcg ccacaacatc gaagacgggt
     1381 cggttcaact cgctgatcac taccagcaga acacaccaat tggaggtaag ttttattcca
     1441 taatacaaat atatattgaa caataatgct tttcagatgg accagtcctc ctccctgata
     1501 atcactacct ttccactcaa tccgctctta gcaaggatcc aaatgagaaa agagatcaca
     1561 tggttcttct cgagtttgtc accgccgccg gaatcaccca cggaatggac gagctttaca
     1621 agtaaatgca aaatcctttc aagcattccc ttcttctcta tcactcttct ttctttttgt
     1681 caaaaaattc tctcgctaat ttatttgctt ttttaatgtt attattttat gactttttat
     1741 agtcactgaa aagtttgcat ctgagtgaag tgaatgctat caaaatgtga ttctgtctga
     1801 tgtactttca caatctctct tcaattccat tttgaagtgc tttaaacccg aaaggttgag
     1861 aaaaatgcga gcgctcaaat atttgtattg tgttcgttga gtgacccaac aaaaagagga
     1921 aactttattg tgccgccaag aaaaaagtct cacaggaaac ggagaatctg tgcggcaatg
     1981 tcttatcatt tgaaacgcgc cgtatcgatg ctgacatcat gcggctcccg tgttagtcga
     2041 gaagatgggt acgggccttt attcatccaa cactctacgg cttcttcaag agctagtagg
     2101 gcaccctgta gttggaaagg gattatttcg tatggcgagc ccataccgtc tttcttgcgg
     2161 aagcttaaca cgataggaag ttggatagtt tcgaacgatg gttccatacg gaacgctgtc
     2221 tcaaggatga gtctgacgga gtgtaactcg aaggctaggt ggaggctcag tgatgataag
     2281 tctgcgatgg tggatgcatg tgtcatggtc atagctgttt cctgtgtgaa attgttatcc
     2341 gctcagaggg cacaatccta ttccgcgcta tccgacaatc tccaagacat taggtggagt
     2401 tcagttcggc gtatggcata tgtcgctgga aagaacatgt gagcaaaagg ccagcaaaag
     2461 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2521 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2581 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2641 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2701 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2761 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2821 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2881 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2941 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3001 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3061 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3121 ctattcaaca aagccgccgt cccgtcaagt cagcgtaaat gggtaggggg cttcaaatcg
     3181 tcctcgtgat accaattcgg agcctgcttt tttgtacaaa cttgttgata atggcaattc
     3241 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3301 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3361 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3421 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagagc cacgctcacc
     3481 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3541 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3601 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3661 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3721 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3781 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3841 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3901 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3961 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4021 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4081 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4141 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4201 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4261 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cagatacctg
     4321 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     4381 agggttttcc cagtcacgac gtt
//
