LOCUS       sequence-361781-        4496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP transcriptional reporter cloning vector for mosTI(unc-119). 
            Transgene inserted upstream of gfp(2xNLS) 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 4496)
  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 4496)
  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: pSEM387 - mosTI unc-119 - NNN - gfp - nls - tbb-2
FEATURES             Location/Qualifiers
     source          1..4496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..31
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15..31
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(655..688)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             773..793
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1733..2056
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2415..2431)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2415..2431)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2428..2450)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2603..3191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2683..2702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3308..3332)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3432..4223,4224..4292))
                     /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     4055..4074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4293..4397)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(4496,1..22)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ccagtcacga cgttgtaaaa cgacggccag tcatgcataa tccgcacgca tctggaataa
       61 ggaagtgcca ttccgcctga cctaagtgct tttcgagatc gccaaaccgg agaacgatca
      121 ggagaatgat gagtcgccac aggagtcggc aagatacgtg cgttatagat ttgctccaaa
      181 ctttttgaaa ctcaaaacgg tcggagcaac tgtggaattc aaagtaggag acatcccaat
      241 ccatcatttc cgaatgatcg aacgtcactt cttcaaagat cgccttctga agtgttttga
      301 ctttgaattc ggattctgta ttccgaattc acgaaacaac tgtgaacata tctatgagtt
      361 ccctcaactc tctcaacaac tcagtgagtc attattctaa aaagtacaat tcaaaagact
      421 aatctctttt tcagtggacg acatgatcaa caatccaaac gagactcgtt ctgacagctt
      481 ctatttcgtc gataacaaac tcgtcatgca caacaaagcc gactactcat atgatgcata
      541 aatcttcatt gttgagttta tcttgttgat ttttgaataa attatcaact ctttactttt
      601 taatgggtta tgaaataaat aaacattgaa aactgataaa caacgttcat ctctataact
      661 tcgtatagca tacattatac gaagttattg cctgagacca gctagcaagc ttgcatgcct
      721 gcaggtcgac tctagaggat ccccgggtac cgagctcagg tctctaaaaa tgccaaagaa
      781 gaagagaaag gtgtcaaaag gagaagaact ctttaccgga gtcgtcccaa ttctcgtcga
      841 gctcgacgga gacgtaaacg gacacaaatt ctcggtttcc ggagaaggag aaggaggtaa
      901 gttttacgaa attctgaagc tcacaaaaca tggagtacat ttttcagatg ctacttatgg
      961 aaaactcacc cttaaattca tttgcaccac cggaaagctc ccagtgccgt ggccgaccct
     1021 tgtgactaca ttctgctacg gagttcaatg cttttcccgc tacccagacc acatgaagcg
     1081 tcacgatttc ttcaaatcag ccatgccaga aggatacgtc caggagcgaa caattttctt
     1141 caaggacgat ggaaactaca agactcgtgc tgaagtcaag tttgaaggag atactctcgt
     1201 gaaccgcatt gagctcaagg taagtttata atctatgatc ttaatttggc aatggaaaaa
     1261 gttttttcag ggaatcgact tcaaagaaga tggaaacatt cttggacaca agctcgaata
     1321 caactataac tcgcacaacg tgtatatcat ggccgacaag caaaaaaatg gaatcaaggt
     1381 caacttcaag attcgccaca acatcgaaga cgggtcggtt caactcgctg atcactacca
     1441 gcagaacaca ccaattggag gtaagtttta ttccataata caaatatata ttgaacaata
     1501 atgcttttca gatggaccag tcctcctccc tgataatcac tacctttcca ctcaatccgc
     1561 tcttagcaag gatccaaatg agaaaagaga tcacatggtt cttctcgagt ttgtcaccgc
     1621 cgccggaatc acccacggaa tggacgagct ttacaagtcg agacgccgca aggctaatcc
     1681 aaccaaactt tctgaaaatg ccaagaaact tgccaaggag gtcgagaact aaatgcaaaa
     1741 tcctttcaag cattcccttc ttctctatca ctcttctttc tttttgtcaa aaaattctct
     1801 cgctaattta tttgcttttt taatgttatt attttatgac tttttatagt cactgaaaag
     1861 tttgcatctg agtgaagtga atgctatcaa aatgtgattc tgtctgatgt actttcacaa
     1921 tctctcttca attccatttt gaagtgcttt aaacccgaaa ggttgagaaa aatgcgagcg
     1981 ctcaaatatt tgtattgtgt tcgttgagtg acccaacaaa aagaggaaac tttattgtgc
     2041 cgccaagaaa aaagtctcac aggaaacgga gaatctgtgc ggcaatgtct tatcatttga
     2101 aacgcgccgt atcgatgctg acatcatgcg gctcccgtgt tagtcgagaa gatgggtacg
     2161 ggcctttatt catccaacac tctacggctt cttcaagagc tagtagggca ccctgtagtt
     2221 ggaaagggat tatttcgtat ggcgagccca taccgtcttt cttgcggaag cttaacacga
     2281 taggaagttg gatagtttcg aacgatggtt ccatacggaa cgctgtctca aggatgagtc
     2341 tgacggagtg taactcgaag gctaggtgga ggctcagtga tgataagtct gcgatggtgg
     2401 atgcatgtgt catggtcata gctgtttcct gtgtgaaatt gttatccgct cagagggcac
     2461 aatcctattc cgcgctatcc gacaatctcc aagacattag gtggagttca gttcggcgta
     2521 tggcatatgt cgctggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2581 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2641 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2701 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2761 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2821 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2881 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2941 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3001 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3061 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3121 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcta ttcaacaaag
     3241 ccgccgtccc gtcaagtcag cgtaaatggg tagggggctt caaatcgtcc tcgtgatacc
     3301 aattcggagc ctgctttttt gtacaaactt gttgataatg gcaattcaag gatcttcacc
     3361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3481 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3541 ccatctggcc ccagtgctgc aatgataccg cgagagccac gctcaccggc tccagattta
     3601 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3661 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3721 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3781 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3841 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3901 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3961 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4021 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4081 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4141 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4201 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4261 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4321 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4381 caaatagggg ttccgcgcac atttccccga aaagtgccag atacctgaaa caaaacccat
     4441 cgtacggcca aggaagtctc caataactgt gatccaccac aagcgccagg gttttc
//
