LOCUS       sequence-317748-        3164 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 3164)
  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 3164)
  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: pSEM271 - MCS mosTI NeoR target
FEATURES             Location/Qualifiers
     source          1..3164
                     /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(588..621)
                     /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(1069..1085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1069..1085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1082..1104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1257..1845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1337..1356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1962..1986)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2086..2877,2878..2946))
                     /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     2709..2728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2947..3051)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3150..3164,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3164,1..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 gaagaacatc aaggtttagc accggctgaa cttttcgctc gacttaaagc
      121 ccgtatgcca gatggtgagg accttgtcgt aacccacgga gatgcttgcc ttccaaatat
      181 aatggtcgaa aatggtcgat tcagtggctt cattgattgt ggaagactcg gagtggcgga
      241 ccgatatcag gatatagcgc ttgcgacgcg agatattgcc gaaggtaagt tttcaaaaaa
      301 ttaagattca gttatggggc ttcagactcg caatgggttt tcaggctttc cctatccaaa
      361 acgctcgttc gagtgtttca gttcggtatg tcgaggcgtt accggacgga aaatatttaa
      421 aataaaatcc tcgtcatttc agagttggga ggcgaatggg cagacagatt cctggttctt
      481 tacggaatag ccgccccgga ctctcaacgt attgcttttt atagattatt ggatgagttt
      541 ttctgatctg ttcgttgttc caataaattg ttattgttcc ctatttgata acttcgtata
      601 gcatacatta tacgaagtta ttgcctgaga ccagctagca agcttgcatg cctgcaggtc
      661 gactctagag gatccccggg taccgagctc aggtctctga gctagcggga gctcaggaaa
      721 cggagaatct gtgcggcaat gtcttatcat ttgaaacgcg ccgtatcgat gctgacatca
      781 tgcggctccc gtgttagtcg agaagatggg tacgggcctt tattcatcca acactctacg
      841 gcttcttcaa gagctagtag ggcaccctgt agttggaaag ggattatttc gtatggcgag
      901 cccataccgt ctttcttgcg gaagcttaac acgataggaa gttggatagt ttcgaacgat
      961 ggttccatac ggaacgctgt ctcaaggatg agtctgacgg agtgtaactc gaaggctagg
     1021 tggaggctca gtgatgataa gtctgcgatg gtggatgcat gtgtcatggt catagctgtt
     1081 tcctgtgtga aattgttatc cgctcagagg gcacaatcct attccgcgct atccgacaat
     1141 ctccaagaca ttaggtggag ttcagttcgg cgtatggcat atgtcgctgg aaagaacatg
     1201 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1261 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1321 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1381 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1441 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1501 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1561 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1621 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1681 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1741 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1801 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1861 ttttctacgg ggtctgacgc tctattcaac aaagccgccg tcccgtcaag tcagcgtaaa
     1921 tgggtagggg gcttcaaatc gtcctcgtga taccaattcg gagcctgctt ttttgtacaa
     1981 acttgttgat aatggcaatt caaggatctt cacctagatc cttttaaatt aaaaatgaag
     2041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2221 accgcgagag ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3061 ccgaaaagtg ccagatacct gaaacaaaac ccatcgtacg gccaaggaag tctccaataa
     3121 ctgtgatcca ccacaagcgc cagggttttc ccagtcacga cgtt
//
