LOCUS       sequence-309323-        5097 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Single copy or array insertion by MosTI using split NeoR selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5097)
  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 5097)
  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: pSEM267 - pEXP(mosTI, neoR)[Pmex-5 | ceGFP(PAT...
FEATURES             Location/Qualifiers
     source          1..5097
                     /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(589..622)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    661..681
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1189..1213
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1222..1242
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2151..2222
                     /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(2230..2254)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2259..2582
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2586..2606)
                     /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(3003..3019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3003..3019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3016..3038)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3191..3779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3271..3290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3896..3920)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(4020..4811,4812..4880))
                     /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     4643..4662
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4881..4985)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(5084..5097,1..9)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgtaaaacga cggccagtca tgcataatcc gcacgcatct ggaataagga agtgccattc
       61 cgcctgacct cgaagaacat caaggtttag caccggctga acttttcgct cgacttaaag
      121 cccgtatgcc agatggtgag gaccttgtcg taacccacgg agatgcttgc cttccaaata
      181 taatggtcga aaatggtcga ttcagtggct tcattgattg tggaagactc ggagtggcgg
      241 accgatatca ggatatagcg cttgcgacgc gagatattgc cgaaggtaag ttttcaaaaa
      301 attaagattc agttatgggg cttcagactc gcaatgggtt ttcaggcttt ccctatccaa
      361 aacgctcgtt cgagtgtttc agttcggtat gtcgaggcgt taccggacgg aaaatattta
      421 aaataaaatc ctcgtcattt cagagttggg aggcgaatgg gcagacagat tcctggttct
      481 ttacggaata gccgccccgg actctcaacg tattgctttt tatagattat tggatgagtt
      541 tttctgatct gttcgttgtt ccaataaatt gttattgttc cctatttgat aacttcgtat
      601 agcatacatt atacgaagtt attgccatag catggccttt gcaggattac gccaagctat
      661 caactttgta tagaaaagtt gggttttcac aacggcaaaa tatcagtttt taaaaaatta
      721 aaccataaaa caaataatat aacccaattt ttacatcaaa ccacaagaaa aaaatacatt
      781 tgggcccacg gataaagaaa ttaaaaaaat acatttttta aaggcgcacc gaattaaaat
      841 tcatttgggt cttaccgcgt ataccgtact ccgtttgttt gatcattttt gtcagcgctg
      901 gcggttgttt tttcatttca tttctgcttc aaagacgttt tctcgaataa tttttcgttt
      961 attctctttt ttaaaattaa tttctagccg taaatgttat aaattcaccc atttaacgca
     1021 aatttcatgg taatctcatg gaaaaatgca gtttctttgt taaagaaagc ttaaatagca
     1081 aaaattcccc gactttcccc aaaatcctgc tcgattttcc gttttctcat tgtattctct
     1141 cttaattaat tttatcgata atcaattgaa tgtttcagac agagaatgac aagtttgtac
     1201 aaaaaagcag gctcgggatc cccaaagaag aagcgtaagg tgtccaaggt aacacttagt
     1261 ttttgttgct ctgtttaaaa attaatttta agggagagga gctgttcacc ggagtcgtcc
     1321 caatcctcgt cgagctcgac ggggacgtca acggacacaa gttctccgtg tccggagagg
     1381 gagagggaga tgccacctac ggaaagctca ccctcaagtt catctgcacc accggaaagc
     1441 tcccagtccc atggccaacc ctcgtcacca ccttctgcta cggagtccaa tgcttctccc
     1501 gttacccagg taagttttct tgagaccttt actaactaac taggtctcat agattttcag
     1561 accacatgaa gcgtcacgac ttcttcaagt ccgccatgcc agagggatac gtccaagagc
     1621 gtaccatctt cttcaaggac gacggaaact acaaggtaag ttatgtagag acctttacta
     1681 actaactagg tctcaaacat tttcagaccc gtgccgaggt caagttcgag ggagacaccc
     1741 tcgtcaaccg tatcgagctc aagggaatcg acttcaagga ggacggtaac atcctgggac
     1801 acaagctgga gtacaactac aactcccaca acgtctacat catggccgac aagcaaaaga
     1861 acggaatcaa ggtcaacttc aaggtaagtt ctttagagac ctttactaac taactaggtc
     1921 tcagtaattt tcagatccgt cacaacatcg aggacggatc ggtccaactc gccgaccact
     1981 accaacaaaa caccccaatc ggtgacggac cagtcctcct cccagacaac cactacctct
     2041 ccacccaatc cgccctctcc aaggacccaa acgagaagcg tgaccacatg gtcctcctcg
     2101 agttcgtcac cgccgccgga atcacccacg gaatggacga gctctacaag tcccgtcgtc
     2161 gtaaggccaa cccaaccaag ctctccgaga acgccaagaa gctcgccaag gaggtcgaga
     2221 acggatccaa cccagctttc ttgtacaaag tgggataaat gcaaaatcct ttcaagcatt
     2281 cccttcttct ctatcactct tctttctttt tgtcaaaaaa ttctctcgct aatttatttg
     2341 cttttttaat gttattattt tatgactttt tatagtcact gaaaagtttg catctgagtg
     2401 aagtgaatgc tatcaaaatg tgattctgtc tgatgtactt tcacaatctc tcttcaattc
     2461 cattttgaag tgctttaaac ccgaaaggtt gagaaaaatg cgagcgctca aatatttgta
     2521 ttgtgttcgt tgagtgaccc aacaaaaaga ggaaacttta ttgtgccgcc aagaaaaaag
     2581 tctcacaact ttattataca tagttgataa ttccctctga cacatgcagc tcgagctagc
     2641 gggagctcag gaaacggaga atctgtgcgg caatgtctta tcatttgaaa cgcgccgtat
     2701 cgatgctgac atcatgcggc tcccgtgtta gtcgagaaga tgggtacggg cctttattca
     2761 tccaacactc tacggcttct tcaagagcta gtagggcacc ctgtagttgg aaagggatta
     2821 tttcgtatgg cgagcccata ccgtctttct tgcggaagct taacacgata ggaagttgga
     2881 tagtttcgaa cgatggttcc atacggaacg ctgtctcaag gatgagtctg acggagtgta
     2941 actcgaaggc taggtggagg ctcagtgatg ataagtctgc gatggtggat gcatgtgtca
     3001 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa tcctattccg
     3061 cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg gcatatgtcg
     3121 ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3181 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3241 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3301 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3361 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3421 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3481 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3541 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3601 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3661 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3721 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3781 aagatccttt gatcttttct acggggtctg acgctctatt caacaaagcc gccgtcccgt
     3841 caagtcagcg taaatgggta gggggcttca aatcgtcctc gtgataccaa ttcggagcct
     3901 gcttttttgt acaaacttgt tgataatggc aattcaagga tcttcaccta gatcctttta
     3961 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4021 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4081 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4141 agtgctgcaa tgataccgcg agagccacgc tcaccggctc cagatttatc agcaataaac
     4201 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4261 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4321 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4381 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4441 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4501 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4561 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4621 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4681 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4741 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4801 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4861 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4921 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4981 ccgcgcacat ttccccgaaa agtgccagat acctgaaaca aaacccatcg tacggccaag
     5041 gaagtctcca ataactgtga tccaccacaa gcgccagggt tttcccagtc acgacgt
//
