LOCUS       sequence-317456-        2522 bp    DNA     circular SYN 15-DEC-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 2522)
  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 2522)
  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: pSEM253 - sgRNA mosTI hygroR
FEATURES             Location/Qualifiers
     source          1..2522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(427..443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(427..443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(440..462)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(615..1203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(695..714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1320..1344)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1444..2235,2236..2304))
                     /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     2067..2086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2305..2409)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(2508..2522,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2522,1..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 gcctccccgc acactcaaca cagcgagcgg cggaacccgc ggggtgtcgg
      121 ccgctcggac acgctcccgt atataacgac gcggcgcgcg tcaagttgtg agtgggagct
      181 tacatatggg tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc
      241 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt gttttttgcc cattttttgt
      301 agtttgagac tggaaaaatc gtaaatcaac catagtttta gggcaaaatt ctcgaaattt
      361 ttctaaaaat aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt
      421 gtcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat
      481 tccgcgctat ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat
      541 gtcgctggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1201 caagaagatc ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc
     1261 ccgtcaagtc agcgtaaatg ggtagggggc ttcaaatcgt cctcgtgata ccaattcgga
     1321 gcctgctttt ttgtacaaac ttgttgataa tggcaattca aggatcttca cctagatcct
     1381 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1441 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1501 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1561 ccccagtgct gcaatgatac cgcgagagcc acgctcaccg gctccagatt tatcagcaat
     1621 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1681 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1741 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1801 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1861 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1921 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1981 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2041 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2101 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2161 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2221 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2281 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2341 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2401 ggttccgcgc acatttcccc gaaaagtgcc agatacctga aacaaaaccc atcgtacggc
     2461 caaggaagtc tccaataact gtgatccacc acaagcgcca gggttttccc agtcacgacg
     2521 tt
//
