LOCUS       sequence-361775-        4647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP co-expression cloning vector for mosTI(unc-119). Transgene 
            inserted upstream of gpd-2 operon::gfp::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 4647)
  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 4647)
  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: pSEM396 - mosTI unc-119 - NNN - operon - gfp -...
FEATURES             Location/Qualifiers
     source          1..4647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..27
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11..27
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(651..684)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      1880..2203
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2562..2578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2562..2578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2575..2597)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2750..3338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2830..2849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3455..3479)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3579..4370,4371..4439))
                     /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     4202..4221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4440..4544)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(4643..4647,1..18)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcacgacgtt gtaaaacgac ggccagtcat gcataatccg cacgcatctg gaataaggaa
       61 gtgccattcc gcctgaccta agtgcttttc gagatcgcca aaccggagaa cgatcaggag
      121 aatgatgagt cgccacagga gtcggcaaga tacgtgcgtt atagatttgc tccaaacttt
      181 ttgaaactca aaacggtcgg agcaactgtg gaattcaaag taggagacat cccaatccat
      241 catttccgaa tgatcgaacg tcacttcttc aaagatcgcc ttctgaagtg ttttgacttt
      301 gaattcggat tctgtattcc gaattcacga aacaactgtg aacatatcta tgagttccct
      361 caactctctc aacaactcag tgagtcatta ttctaaaaag tacaattcaa aagactaatc
      421 tctttttcag tggacgacat gatcaacaat ccaaacgaga ctcgttctga cagcttctat
      481 ttcgtcgata acaaactcgt catgcacaac aaagccgact actcatatga tgcataaatc
      541 ttcattgttg agtttatctt gttgattttt gaataaatta tcaactcttt actttttaat
      601 gggttatgaa ataaataaac attgaaaact gataaacaac gttcatctct ataacttcgt
      661 atagcataca ttatacgaag ttattgcctg agaccagcta gcaagcttgc atgcctgcag
      721 gtcgactcta gaggatcccc gggtaccgag ctcaggtctc ttgagctgtc tcatcctact
      781 ttcacctagt taactgcttg tcttaaaatc tatgcttctc tttagtatct aaaattttcc
      841 tagaagctta caagtatata aattgtctct tctcaataaa ggttgtatat ttattcatct
      901 tattgaatct gccatttcct cgtttttgcg agtttatata ccttccaatt ttctttctat
      961 tgtattttca acttctaatt ttaattcagg gaaactgctt caacgcatca tgtcaaaagg
     1021 agaagaactc tttaccggag tcgtcccaat tctcgtcgag ctcgacggag acgtaaacgg
     1081 acacaaattc tcggtttccg gagaaggaga aggaggtaag ttttacgaaa ttctgaagct
     1141 cacaaaacat ggagtacatt tttcagatgc tacttatgga aaactcaccc ttaaattcat
     1201 ttgcaccacc ggaaagctcc cagtgccgtg gccgaccctt gtgactacat tctgctacgg
     1261 agttcaatgc ttttcccgct acccagacca catgaagcgt cacgatttct tcaaatcagc
     1321 catgccagaa ggatacgtcc aggagcgaac aattttcttc aaggacgatg gaaactacaa
     1381 gactcgtgct gaagtcaagt ttgaaggaga tactctcgtg aaccgcattg agctcaaggt
     1441 aagtttataa tctatgatct taatttggca atggaaaaag ttttttcagg gaatcgactt
     1501 caaagaagat ggaaacattc ttggacacaa gctcgaatac aactataact cgcacaacgt
     1561 gtatatcatg gccgacaagc aaaaaaatgg aatcaaggtc aacttcaaga ttcgccacaa
     1621 catcgaagac gggtcggttc aactcgctga tcactaccag cagaacacac caattggagg
     1681 taagttttat tccataatac aaatatatat tgaacaataa tgcttttcag atggaccagt
     1741 cctcctccct gataatcact acctttccac tcaatccgct cttagcaagg atccaaatga
     1801 gaaaagagat cacatggttc ttctcgagtt tgtcaccgcc gccggaatca cccacggaat
     1861 ggacgagctt tacaagtaaa tgcaaaatcc tttcaagcat tcccttcttc tctatcactc
     1921 ttctttcttt ttgtcaaaaa attctctcgc taatttattt gcttttttaa tgttattatt
     1981 ttatgacttt ttatagtcac tgaaaagttt gcatctgagt gaagtgaatg ctatcaaaat
     2041 gtgattctgt ctgatgtact ttcacaatct ctcttcaatt ccattttgaa gtgctttaaa
     2101 cccgaaaggt tgagaaaaat gcgagcgctc aaatatttgt attgtgttcg ttgagtgacc
     2161 caacaaaaag aggaaacttt attgtgccgc caagaaaaaa gtctcacagg aaacggagaa
     2221 tctgtgcggc aatgtcttat catttgaaac gcgccgtatc gatgctgaca tcatgcggct
     2281 cccgtgttag tcgagaagat gggtacgggc ctttattcat ccaacactct acggcttctt
     2341 caagagctag tagggcaccc tgtagttgga aagggattat ttcgtatggc gagcccatac
     2401 cgtctttctt gcggaagctt aacacgatag gaagttggat agtttcgaac gatggttcca
     2461 tacggaacgc tgtctcaagg atgagtctga cggagtgtaa ctcgaaggct aggtggaggc
     2521 tcagtgatga taagtctgcg atggtggatg catgtgtcat ggtcatagct gtttcctgtg
     2581 tgaaattgtt atccgctcag agggcacaat cctattccgc gctatccgac aatctccaag
     2641 acattaggtg gagttcagtt cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa
     2701 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2761 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2821 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2881 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2941 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3001 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3061 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3121 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3181 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3241 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3301 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3361 cggggtctga cgctctattc aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag
     3421 ggggcttcaa atcgtcctcg tgataccaat tcggagcctg cttttttgta caaacttgtt
     3481 gataatggca attcaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3541 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3601 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3661 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3721 gagccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3781 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3841 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3901 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3961 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4021 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4081 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4141 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4201 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4261 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4321 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4381 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4441 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4501 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4561 gtgccagata cctgaaacaa aacccatcgt acggccaagg aagtctccaa taactgtgat
     4621 ccaccacaag cgccagggtt ttcccag
//
