LOCUS       sequence-309319-        5083 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 5083)
  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 5083)
  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: pSEM266 - pEXP(mosTI, hygroR)[Pmex-5 | ceGFP(P...
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..18
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2..18
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(575..608)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    647..667
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1175..1199
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1208..1228
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2137..2208
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(2216..2240)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2245..2568
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2572..2592)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2989..3005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2989..3005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3002..3024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3177..3765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3257..3276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3882..3906)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(4006..4797,4798..4866))
                     /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     4629..4648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4867..4971)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(5070..5083,1..9)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgtaaaacga cggccagtca tgcataatcc gcacgcatct ggaataagga agtgccattc
       61 cgcctgacct atgcatggaa caacaaacca gatattttga acgacgacac ccagaattag
      121 ccggatctcc taggcttcga gcttacatgc ttagaatcgg attagatcag ttgtaccaat
      181 ctcttgtgga tggtaacttt gacgacgctg cgtgggctca agggcgttgt gatgccattg
      241 tgcgttcagg ggcaggcacg gtaggacgaa ctcagattgc aagaaggtaa gttttcaaaa
      301 aattaagatt cagttatggg aagtgggagc ttacatattt tcaggctttc cctatccaaa
      361 acgctcgttc gagtgtttca gttcggtatg tcgaggcgtt accggacgga aaatatttaa
      421 aataaaatcc tcgtcatttc aggtcagctg cggtttggac tgacggatgc gtcgaggtcc
      481 ttgctgactc aggtaataga agaccttcta caagaccacg tgccaaagag taatctgttc
      541 gttgttccaa taaattgtta ttgttcccta tttgataact tcgtatagca tacattatac
      601 gaagttattg ccatagcatg gcctttgcag gattacgcca agctatcaac tttgtataga
      661 aaagttgggt tttcacaacg gcaaaatatc agtttttaaa aaattaaacc ataaaacaaa
      721 taatataacc caatttttac atcaaaccac aagaaaaaaa tacatttggg cccacggata
      781 aagaaattaa aaaaatacat tttttaaagg cgcaccgaat taaaattcat ttgggtctta
      841 ccgcgtatac cgtactccgt ttgtttgatc atttttgtca gcgctggcgg ttgttttttc
      901 atttcatttc tgcttcaaag acgttttctc gaataatttt tcgtttattc tcttttttaa
      961 aattaatttc tagccgtaaa tgttataaat tcacccattt aacgcaaatt tcatggtaat
     1021 ctcatggaaa aatgcagttt ctttgttaaa gaaagcttaa atagcaaaaa ttccccgact
     1081 ttccccaaaa tcctgctcga ttttccgttt tctcattgta ttctctctta attaatttta
     1141 tcgataatca attgaatgtt tcagacagag aatgacaagt ttgtacaaaa aagcaggctc
     1201 gggatcccca aagaagaagc gtaaggtgtc caaggtaaca cttagttttt gttgctctgt
     1261 ttaaaaatta attttaaggg agaggagctg ttcaccggag tcgtcccaat cctcgtcgag
     1321 ctcgacgggg acgtcaacgg acacaagttc tccgtgtccg gagagggaga gggagatgcc
     1381 acctacggaa agctcaccct caagttcatc tgcaccaccg gaaagctccc agtcccatgg
     1441 ccaaccctcg tcaccacctt ctgctacgga gtccaatgct tctcccgtta cccaggtaag
     1501 ttttcttgag acctttacta actaactagg tctcatagat tttcagacca catgaagcgt
     1561 cacgacttct tcaagtccgc catgccagag ggatacgtcc aagagcgtac catcttcttc
     1621 aaggacgacg gaaactacaa ggtaagttat gtagagacct ttactaacta actaggtctc
     1681 aaacattttc agacccgtgc cgaggtcaag ttcgagggag acaccctcgt caaccgtatc
     1741 gagctcaagg gaatcgactt caaggaggac ggtaacatcc tgggacacaa gctggagtac
     1801 aactacaact cccacaacgt ctacatcatg gccgacaagc aaaagaacgg aatcaaggtc
     1861 aacttcaagg taagttcttt agagaccttt actaactaac taggtctcag taattttcag
     1921 atccgtcaca acatcgagga cggatcggtc caactcgccg accactacca acaaaacacc
     1981 ccaatcggtg acggaccagt cctcctccca gacaaccact acctctccac ccaatccgcc
     2041 ctctccaagg acccaaacga gaagcgtgac cacatggtcc tcctcgagtt cgtcaccgcc
     2101 gccggaatca cccacggaat ggacgagctc tacaagtccc gtcgtcgtaa ggccaaccca
     2161 accaagctct ccgagaacgc caagaagctc gccaaggagg tcgagaacgg atccaaccca
     2221 gctttcttgt acaaagtggg ataaatgcaa aatcctttca agcattccct tcttctctat
     2281 cactcttctt tctttttgtc aaaaaattct ctcgctaatt tatttgcttt tttaatgtta
     2341 ttattttatg actttttata gtcactgaaa agtttgcatc tgagtgaagt gaatgctatc
     2401 aaaatgtgat tctgtctgat gtactttcac aatctctctt caattccatt ttgaagtgct
     2461 ttaaacccga aaggttgaga aaaatgcgag cgctcaaata tttgtattgt gttcgttgag
     2521 tgacccaaca aaaagaggaa actttattgt gccgccaaga aaaaagtctc acaactttat
     2581 tatacatagt tgataattcc ctctgacaca tgcagctcga gctggcggga gctcaggaaa
     2641 cggagaatct gtgcggcaat gtcttatcat ttgaaacgcg ccgtatcgat gctgacatca
     2701 tgcggctccc gtgttagtcg agaagatggg tacgggcctt tattcatcca acactctacg
     2761 gcttcttcaa gagctagtag ggcaccctgt agttggaaag ggattatttc gtatggcgag
     2821 cccataccgt ctttcttgcg gaagcttaac acgataggaa gttggatagt ttcgaacgat
     2881 ggttccatac ggaacgctgt ctcaaggatg agtctgacgg agtgtaactc gaaggctagg
     2941 tggaggctca gtgatgataa gtctgcgatg gtggatgcat gtgtcatggt catagctgtt
     3001 tcctgtgtga aattgttatc cgctcagagg gcacaatcct attccgcgct atccgacaat
     3061 ctccaagaca ttaggtggag ttcagttcgg cgtatggcat atgtcgctgg aaagaacatg
     3121 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3181 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3241 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3301 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3361 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3421 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3481 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3541 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3601 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3661 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3721 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3781 ttttctacgg ggtctgacgc tctattcaac aaagccgccg tcccgtcaag tcagcgtaaa
     3841 tgggtagggg gcttcaaatc gtcctcgtga taccaattcg gagcctgctt ttttgtacaa
     3901 acttgttgat aatggcaatt caaggatctt cacctagatc cttttaaatt aaaaatgaag
     3961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4141 accgcgagag ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4861 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4981 ccgaaaagtg ccagatacct gaaacaaaac ccatcgtacg gccaaggaag tctccaataa
     5041 ctgtgatcca ccacaagcgc cagggttttc ccagtcacga cgt
//
