LOCUS       sequence-361685-        4740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear GFP co-expression cloning vector for mosTI(unc-119). 
            Transgene inserted upstream of gpd-2 operon::gfp(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 4740)
  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 4740)
  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: pSEM397 - mosTI unc-119 - NNN - operon - gfp -...
FEATURES             Location/Qualifiers
     source          1..4740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..26
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10..26
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(650..683)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1012..1032
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1972..2295
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2654..2670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2654..2670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2667..2689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2842..3430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2922..2941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3547..3571)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3671..4462,4463..4531))
                     /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     4294..4313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4532..4636)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(4735..4740,1..17)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cacgacgttg taaaacgacg gccagtcatg cataatccgc acgcatctgg aataaggaag
       61 tgccattccg cctgacctaa gtgcttttcg agatcgccaa accggagaac gatcaggaga
      121 atgatgagtc gccacaggag tcggcaagat acgtgcgtta tagatttgct ccaaactttt
      181 tgaaactcaa aacggtcgga gcaactgtgg aattcaaagt aggagacatc ccaatccatc
      241 atttccgaat gatcgaacgt cacttcttca aagatcgcct tctgaagtgt tttgactttg
      301 aattcggatt ctgtattccg aattcacgaa acaactgtga acatatctat gagttccctc
      361 aactctctca acaactcagt gagtcattat tctaaaaagt acaattcaaa agactaatct
      421 ctttttcagt ggacgacatg atcaacaatc caaacgagac tcgttctgac agcttctatt
      481 tcgtcgataa caaactcgtc atgcacaaca aagccgacta ctcatatgat gcataaatct
      541 tcattgttga gtttatcttg ttgatttttg aataaattat caactcttta ctttttaatg
      601 ggttatgaaa taaataaaca ttgaaaactg ataaacaacg ttcatctcta taacttcgta
      661 tagcatacat tatacgaagt tattgcctga gaccagctag caagcttgca tgcctgcagg
      721 tcgactctag aggatccccg ggtaccgagc tcaggtctct tgagctgtct catcctactt
      781 tcacctagtt aactgcttgt cttaaaatct atgcttctct ttagtatcta aaattttcct
      841 agaagcttac aagtatataa attgtctctt ctcaataaag gttgtatatt tattcatctt
      901 attgaatctg ccatttcctc gtttttgcga gtttatatac cttccaattt tctttctatt
      961 gtattttcaa cttctaattt taattcaggg aaactgcttc aacgcatcat gccaaagaag
     1021 aagagaaagg tgtcaaaagg agaagaactc tttaccggag tcgtcccaat tctcgtcgag
     1081 ctcgacggag acgtaaacgg acacaaattc tcggtttccg gagaaggaga aggaggtaag
     1141 ttttacgaaa ttctgaagct cacaaaacat ggagtacatt tttcagatgc tacttatgga
     1201 aaactcaccc ttaaattcat ttgcaccacc ggaaagctcc cagtgccgtg gccgaccctt
     1261 gtgactacat tctgctacgg agttcaatgc ttttcccgct acccagacca catgaagcgt
     1321 cacgatttct tcaaatcagc catgccagaa ggatacgtcc aggagcgaac aattttcttc
     1381 aaggacgatg gaaactacaa gactcgtgct gaagtcaagt ttgaaggaga tactctcgtg
     1441 aaccgcattg agctcaaggt aagtttataa tctatgatct taatttggca atggaaaaag
     1501 ttttttcagg gaatcgactt caaagaagat ggaaacattc ttggacacaa gctcgaatac
     1561 aactataact cgcacaacgt gtatatcatg gccgacaagc aaaaaaatgg aatcaaggtc
     1621 aacttcaaga ttcgccacaa catcgaagac gggtcggttc aactcgctga tcactaccag
     1681 cagaacacac caattggagg taagttttat tccataatac aaatatatat tgaacaataa
     1741 tgcttttcag atggaccagt cctcctccct gataatcact acctttccac tcaatccgct
     1801 cttagcaagg atccaaatga gaaaagagat cacatggttc ttctcgagtt tgtcaccgcc
     1861 gccggaatca cccacggaat ggacgagctt tacaagtcga gacgccgcaa ggctaatcca
     1921 accaaacttt ctgaaaatgc caagaaactt gccaaggagg tcgagaacta aatgcaaaat
     1981 cctttcaagc attcccttct tctctatcac tcttctttct ttttgtcaaa aaattctctc
     2041 gctaatttat ttgctttttt aatgttatta ttttatgact ttttatagtc actgaaaagt
     2101 ttgcatctga gtgaagtgaa tgctatcaaa atgtgattct gtctgatgta ctttcacaat
     2161 ctctcttcaa ttccattttg aagtgcttta aacccgaaag gttgagaaaa atgcgagcgc
     2221 tcaaatattt gtattgtgtt cgttgagtga cccaacaaaa agaggaaact ttattgtgcc
     2281 gccaagaaaa aagtctcaca ggaaacggag aatctgtgcg gcaatgtctt atcatttgaa
     2341 acgcgccgta tcgatgctga catcatgcgg ctcccgtgtt agtcgagaag atgggtacgg
     2401 gcctttattc atccaacact ctacggcttc ttcaagagct agtagggcac cctgtagttg
     2461 gaaagggatt atttcgtatg gcgagcccat accgtctttc ttgcggaagc ttaacacgat
     2521 aggaagttgg atagtttcga acgatggttc catacggaac gctgtctcaa ggatgagtct
     2581 gacggagtgt aactcgaagg ctaggtggag gctcagtgat gataagtctg cgatggtgga
     2641 tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc agagggcaca
     2701 atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag ttcggcgtat
     2761 ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3241 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat tcaacaaagc
     3481 cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtcct cgtgatacca
     3541 attcggagcc tgcttttttg tacaaacttg ttgataatgg caattcaagg atcttcacct
     3601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3781 catctggccc cagtgctgca atgataccgc gagagccacg ctcaccggct ccagatttat
     3841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4501 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4561 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4621 aaataggggt tccgcgcaca tttccccgaa aagtgccaga tacctgaaac aaaacccatc
     4681 gtacggccaa ggaagtctcc aataactgtg atccaccaca agcgccaggg ttttcccagt
//
