LOCUS       sequence-309322-        2534 bp    DNA     circular SYN 15-DEC-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 2534)
  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 2534)
  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: pSEM322 - array co-injection fragment mosTI neoR
FEATURES             Location/Qualifiers
     source          1..2534
                     /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(349..382)
                     /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(439..455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(439..455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(452..474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(627..1215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(707..726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1332..1356)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1456..2247,2248..2316))
                     /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     2079..2098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2317..2421)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(2520..2534,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2534,1..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 cgctacattg cgagtctgaa gcttttcagg ctttccctat ccaaaacgct
      121 cgttcgagtg tttcagttcg gtatgtcgag gcgttaccgg acggaaaata tttaaaataa
      181 aatcctcgtc atttcagagt tgggaggcga atgggcagac agattcctgg ttctttacgg
      241 aatagccgcc ccggactctc aacgtattgc tttttataga ttattggatg agtttttctg
      301 atctgttcgt tgttccaata aattgttatt gttccctatt tggctagcat aacttcgtat
      361 agcatacatt atacgaagtt ataggctagg tggaggctca gtgatgataa gtctgcgatg
      421 gtggatgcat gtgtcatggt catagctgtt tcctgtgtga aattgttatc cgctcagagg
      481 gcacaatcct attccgcgct atccgacaat ctccaagaca ttaggtggag ttcagttcgg
      541 cgtatggcat atgtcgctgg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
      841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
      961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tctattcaac
     1261 aaagccgccg tcccgtcaag tcagcgtaaa tgggtagggg gcttcaaatc gtcctcgtga
     1321 taccaattcg gagcctgctt ttttgtacaa acttgttgat aatggcaatt caaggatctt
     1381 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1441 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1501 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1561 cttaccatct ggccccagtg ctgcaatgat accgcgagag ccacgctcac cggctccaga
     1621 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1681 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1741 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1801 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1861 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1921 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1981 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2041 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2101 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2161 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2221 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2281 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2341 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2401 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccagatacct gaaacaaaac
     2461 ccatcgtacg gccaaggaag tctccaataa ctgtgatcca ccacaagcgc cagggttttc
     2521 ccagtcacga cgtt
//
