LOCUS       sequence-309318-        2521 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Single copy or array insertion by MosTI using split hygroR 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2521)
  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 2521)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pSEM321 - array co-injection fragment mosTI hy...
FEATURES             Location/Qualifiers
     source          1..2521
                     /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(336..369)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(426..442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(426..442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(439..461)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(614..1202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(694..713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1319..1343)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1443..2234,2235..2303))
                     /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     2066..2085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2304..2408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(2507..2521,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2521,1..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 gcctgacctc cgccatatgt aagctcccac ttttttcagg ctttccctat ccaaaacgct
      121 cgttcgagtg tttcagttcg gtatgtcgag gcgttaccgg acggaaaata tttaaaataa
      181 aatcctcgtc atttcaggtc agctgcggtt tggactgacg gatgcgtcga ggtccttgct
      241 gactcaggta atagaagacc ttctacaaga ccacgtgcca aagagtaatc tgttcgttgt
      301 tccaataaat tgttattgtt ccctatttgg ctagcataac ttcgtatagc atacattata
      361 cgaagttata ggctaggtgg aggctcagtg atgataagtc tgcgatggtg gatgcatgtg
      421 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcagagggca caatcctatt
      481 ccgcgctatc cgacaatctc caagacatta ggtggagttc agttcggcgt atggcatatg
      541 tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1021 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1201 aagaagatcc tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc
     1261 cgtcaagtca gcgtaaatgg gtagggggct tcaaatcgtc ctcgtgatac caattcggag
     1321 cctgcttttt tgtacaaact tgttgataat ggcaattcaa ggatcttcac ctagatcctt
     1381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1561 cccagtgctg caatgatacc gcgagagcca cgctcaccgg ctccagattt atcagcaata
     1621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2401 gttccgcgca catttccccg aaaagtgcca gatacctgaa acaaaaccca tcgtacggcc
     2461 aaggaagtct ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt
     2521 t
//
