LOCUS       sequence-309324-        3209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single copy or array insertion by MosTI using split Pmlc-2 GFP 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3209)
  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 3209)
  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: pSEM272 - MCS mosTI Pmlc-2_GFP target
FEATURES             Location/Qualifiers
     source          1..3209
                     /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(627..660)
                     /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(1114..1130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1114..1130)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1127..1149)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1302..1890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1382..1401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2007..2031)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2131..2922,2923..2991))
                     /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     2754..2773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2992..3096)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3195..3209,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3209,1..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 gcctgaccta actataattc ccataatgtc tatatcatgg ccgacaagca aaagaatgga
      121 atcaaagtca acttcaagat aagacataat atagaggacg gctctgtcca gctggccgac
      181 cattaccagc agaacacccc gattggagat ggtccggtcc ttcttccaga caatcactat
      241 cttagtacgc aaagtgctct ctcgaaaggt aagttttcaa aaaattaaga ttcagttatg
      301 ggccagacac atcctattct tttcaggctt tccctatcca aaacgctcgt tcgagtgttt
      361 cagttcggta tgtcgaggcg ttaccggacg gaaaatattt aaaataaaat cctcgtcatt
      421 tcagatccta acgagaagcg tgaccatatg gttctcttag agtttgtaac cgccgctggc
      481 ataacacacg gtatggatga gctgtacaaa tcacgaagaa gaaaggctaa ccctactaaa
      541 ttgtcggaga acgcaaaaaa gttggcgaaa gaagttgaga actagtctgt tcgttgttcc
      601 aataaattgt tattgttccc tatttgataa cttcgtatag catacattat acgaagttat
      661 tgcctgagac cagctagcaa gcttgcatgc ctgcaggtcg actctagagg atccccgggt
      721 accgagctca ggtctctgag ctggcgggag cacgtggtca ggaaacggag aatctgtgcg
      781 gcaatgtctt atcatttgaa acgcgccgta tcgatgctga catcatgcgg ctcccgtgtt
      841 agtcgagaag atgggtacgg gcctttattc atccaacact ctacggcttc ttcaagagct
      901 agtagggcac cctgtagttg gaaagggatt atttcgtatg gcgagcccat accgtctttc
      961 ttgcggaagc ttaacacgat aggaagttgg atagtttcga acgatggttc catacggaac
     1021 gctgtctcaa ggatgagtct gacggagtgt aactcgaagg ctaggtggag gctcagtgat
     1081 gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg
     1141 ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg
     1201 tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag
     1261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1741 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1861 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1921 gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc
     1981 aaatcgtcct cgtgatacca attcggagcc tgcttttttg tacaaacttg ttgataatgg
     2041 caattcaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2101 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2161 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2221 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg
     2281 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2341 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2401 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2461 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2521 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2581 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2641 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2701 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2761 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2821 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2881 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2941 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3061 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccaga
     3121 tacctgaaac aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca
     3181 agcgccaggg ttttcccagt cacgacgtt
//
