LOCUS       sequence-309316-        3150 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 3150)
  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 3150)
  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: pSEM270 - MCS mosTI hygroR target
FEATURES             Location/Qualifiers
     source          1..3150
                     /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(574..607)
                     /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(1055..1071)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1055..1071)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1068..1090)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1243..1831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1323..1342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1948..1972)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2072..2863,2864..2932))
                     /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     2695..2714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2933..3037)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3136..3150,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3150,1..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 tgcatggaac aacaaaccag atattttgaa cgacgacacc cagaattagc
      121 cggatctcct aggcttcgag cttacatgct tagaatcgga ttagatcagt tgtaccaatc
      181 tcttgtggat ggtaactttg acgacgctgc gtgggctcaa gggcgttgtg atgccattgt
      241 gcgttcaggg gcaggcacgg taggacgaac tcagattgca agaaggtaag ttttcaaaaa
      301 attaagattc agttatggga agtgggagct tacatatttt caggctttcc ctatccaaaa
      361 cgctcgttcg agtgtttcag ttcggtatgt cgaggcgtta ccggacggaa aatatttaaa
      421 ataaaatcct cgtcatttca ggtcagctgc ggtttggact gacggatgcg tcgaggtcct
      481 tgctgactca ggtaatagaa gaccttctac aagaccacgt gccaaagagt aatctgttcg
      541 ttgttccaat aaattgttat tgttccctat ttgataactt cgtatagcat acattatacg
      601 aagttattgc ctgagaccag ctagcaagct tgcatgcctg caggtcgact ctagaggatc
      661 cccgggtacc gagctcaggt ctctgagctg gcgggagctc aggaaacgga gaatctgtgc
      721 ggcaatgtct tatcatttga aacgcgccgt atcgatgctg acatcatgcg gctcccgtgt
      781 tagtcgagaa gatgggtacg ggcctttatt catccaacac tctacggctt cttcaagagc
      841 tagtagggca ccctgtagtt ggaaagggat tatttcgtat ggcgagccca taccgtcttt
      901 cttgcggaag cttaacacga taggaagttg gatagtttcg aacgatggtt ccatacggaa
      961 cgctgtctca aggatgagtc tgacggagtg taactcgaag gctaggtgga ggctcagtga
     1021 tgataagtct gcgatggtgg atgcatgtgt catggtcata gctgtttcct gtgtgaaatt
     1081 gttatccgct cagagggcac aatcctattc cgcgctatcc gacaatctcc aagacattag
     1141 gtggagttca gttcggcgta tggcatatgt cgctggaaag aacatgtgag caaaaggcca
     1201 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1261 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1321 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1381 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1441 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1501 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1561 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1621 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1681 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1861 tgacgctcta ttcaacaaag ccgccgtccc gtcaagtcag cgtaaatggg tagggggctt
     1921 caaatcgtcc tcgtgatacc aattcggagc ctgctttttt gtacaaactt gttgataatg
     1981 gcaattcaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2041 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2101 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2161 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagagccac
     2221 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2281 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2341 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2401 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2461 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2521 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2581 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2641 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2701 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2761 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2821 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2881 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2941 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3001 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccag
     3061 atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt gatccaccac
     3121 aagcgccagg gttttcccag tcacgacgtt
//
