LOCUS       sequence-250732-        7386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  modified pCFJ151 for C. elegans mosSCI at ttTi5605 or compatible 
            sites; floxed C. br. unc-119(+) selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7386)
  AUTHORS   Aram R, MacGillivray K, Li C, Saltzman A
  TITLE     Tools for Mos1-mediated single copy insertion (mosSCI) with 
            excisable unc-119(+) or NeoR (G418) selection cassettes.
  JOURNAL   MicroPubl Biol. 2019 Aug 27;2019. doi: 
            10.17912/micropub.biology.000146.
  PUBMED    32550440
REFERENCE   2  (bases 1 to 7386)
  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: "MicroPubl 
            Biol."; date: "2019-08-27"; volume: "2019. doi"; pages: " 
            10.17912/micropub.biology.000146"
COMMENT     Sequence Label: pALS186 (ttTi5605-MCS loxP::C.br.unc-119(+)::l...
FEATURES             Location/Qualifiers
     source          1..7386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1408..1441)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1590..1601)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(3570..3603)
                     /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(5252..5271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5371..5393
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5431..5449)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5517..5621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5622..5690,5691..6482)
                     /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     complement(5840..5859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6653..7241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7142..7161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggccctgtc ggattatggg ctcttctttt atcctctact ttctttctag aaaatgcaaa
       61 tgttgagtca gaaattgctg aatggagtac aactgcaatg ggtttagctt atgtttctgg
      121 aacaattact ggtggaattg tgattgaaaa gtaaggttat atttcatttc aataaatgtt
      181 accacagtat aacaaaaaca agttacattg ttcagagtat gtctgaatgt attattatta
      241 ttgtatttgt gcatcagtac tgtagttgta ctgaagaagg gtttaaaata atctcaacgt
      301 ttttaagacg agttagtgag aatacggtcg cagttaggat gtttggacat agttttatta
      361 gataatcgtg aactaactca ctataaaaac ccgctaactc attgaataag atagttttaa
      421 acgttcaaaa tttgaagaca aaattgtatg aataagttta cagagttggt cgacgcaagc
      481 ttctccttct cttcacattt ctcaataatt tagctctaat tgcattcgta ttcttcgcaa
      541 aaatacgtat tctgattgac ccgatgaaat acggatgcct agttgcattg atcatctacg
      601 gttatactta tgggtgaggt ttttcaaata tattttgatt taatcgaata atgaaatttc
      661 agaactggag ttggtccaat ttcgtggttt atatcctctg aactggtacc tcaaaaacat
      721 cgtagtattg ctcaatctgt ggcatattca atgaatactg taatggtggt tatcagtaca
      781 tttacagtat tacctctata ttccgttatt ggtagttatg ctttccttat tctatattca
      841 attccttcat ttataagtat gcttatatta tttcgctact taccggaaac caaaggacga
      901 gagattcatg atattgtgaa tgaacttaaa aaaaaatgaa aacctttgaa aacgttcata
      961 cggccgaaat tttacttgat cgagccaatt gaacttttgt taagacttac agaaaagtga
     1021 gagattttgt tacaaaaaag tgagaaaaaa gcgaaaatga acaggaaaaa gaagaaaaat
     1081 acacccgaag tatcatataa aaattaaaaa gattgtttga cctggcggaa cttttttgca
     1141 aaaactctga atatccatgg cacatgcttc gtgcaaaaca ggatccaaat gaatatcaac
     1201 aagagattct tgaagacgac gagccacttg ctcagagcat tgcttatctc gaatgagacc
     1261 cttgttaaaa tcatttttca aacattatat gacattgtcg aaatgtcctc ctgattccat
     1321 gatggtagca aactcacttc gtcttaaggc cttgactaga gggtaccaga gctcacctag
     1381 gtattcctcg agacgtacgg tgcgcgcata acttcgtata gcatacatta tacgaagtta
     1441 tctttgagcc aatttatcca agtccttgta aaaagtattc gaaaattgtt aacggataaa
     1501 atgtttatta taatatcaaa aacaatttgt cagttgacca ctttttgata tatttgacag
     1561 aaaccgggat gaattggctc aaagttaggg cgcccttcta ttacagggtt tctgataaca
     1621 aaccggttat taactcccaa caagggatga tttcaattca tcatgctcaa attgacccaa
     1681 attaagttac atgacaaatt catcgccctt ttcaactctt cttggctcat catctgttat
     1741 tctgttccta ttctgtctgc accctatacc cctttgcata ctctctcgtc attcctttct
     1801 tggatagtgt cttcttctcc ccagctctgc tacttctatg acttgcgccc gctgcttttc
     1861 cgctcgttcc tctcctctct gacgtcttct tctgcttctt ttctccttcc tttttccctt
     1921 ctcaatcttc ctctttttcc catttctgtc aatcattcga agaagaagaa gaagaagacc
     1981 ctcattcatt tcattttttt ttctgtcggt gtgtgctgct cggttaagag tgagctctct
     2041 attccacgtc ttcttctttt tcttcttcga ttcgaatcaa tcactccacc aaaacgcatt
     2101 cgtttttggg attcaccccg cggttcgcaa caggtttctt tttcaaatta ttagcgttat
     2161 aaatagaaaa atggtggagt ttcaataaaa atgataattt acaaagtgta ttttgattac
     2221 atgtactcaa aaaggttgaa atttttcata ccagttttcc ggaaatccat ctgatatcat
     2281 tatcgtattt ttctttttaa aaatgttttc aaaaaaaaac aaaatatagc tggttatttg
     2341 gcaccctcta attaccattt tcctgtcaca ccacactctt ttccttcttc cacttctttc
     2401 cgttttcagc cgcttccaac caaaccgata tgaaagccga gcaacaacga tcgattccac
     2461 ccggctcggc gaccttcccg tcgcaggtga gactcagaaa actagagaaa ccgctcaact
     2521 aactcttgat atccgatttc attcttttct cttttctttt tgttgaactt ttccatattt
     2581 ccagatgcca cggccaccac caagcaccga acaaggaatc acaacggaat cggagcttgc
     2641 gaagaaagct caaatcactc cgaacgacgt tttagcactt ccgggaatca ctcaaggtat
     2701 cttgctccct ttggatttca gtttaaaaca taaatttagg attcttatgc tccccatctg
     2761 cgaacatcta taacatcgag ttcaccaagt tccaaatccg tgatctggac actgagcaag
     2821 tgcttttcga gatcgccaaa ccggagaacg atcaggagaa tgatgagtcg ccacaggagt
     2881 cggcaagata cgtgcgttat agatttgctc caaacttttt gaaactcaaa acggtcggag
     2941 caactgtgga attcaaagta ggagacatcc caatccatca tttccgaatg atcgaacgtc
     3001 acttcttcaa agatcgcctt ctgaagtgtt ttgactttga attcggattc tgtattccga
     3061 attcacgaaa caactgtgaa catatctatg agttccctca actctctcaa caactcagtg
     3121 agtcattatt ctaaaaagta caattcaaaa gactaatctc tttttcagtg gacgacatga
     3181 tcaacaatcc aaacgagact cgttctgaca gcttctattt cgtcgataac aaactcgtca
     3241 tgcacaacaa agccgactac tcatatgatg cataaatatt taatacaaaa atgttctgga
     3301 taattattct gtcgaataga aaaaaaactc caaatgtgat taaattccaa taattcctgt
     3361 ctagtttgtt ccttcccctt ccccttctca tgttcaatgc attcctaagc ttttcagttc
     3421 cccccttgtt tctatatttt ttcgctgtcc tgtcacactc gctaaaaaca ctaatcacac
     3481 ggaaatctgt tttcaataaa aactccaact ttaactcatt ttcaatttca actgaaagat
     3541 tttttcatta gagaatgtct agaactagga taacttcgta tagcatacat tatacgaagt
     3601 tatagatctg cccactagtg agtcgtatta taagtgcaag taagatcagt gtttgtttcg
     3661 atatcagtct gtttcgtaac ggtcttctgt ataactacaa aaaagaataa aaaaccgtat
     3721 cacaactgag aaaaaagaca aaacttattg aacagtgttc ctttcaaaaa attattttgt
     3781 aaaaaatcga gtttttctcc aaaataacgc acgcaatcaa aaacaagaaa actaacgtgt
     3841 tctcctgggc gtggaactcc ttatcaagat taccattatt gtctgtcaga attgatattt
     3901 caattatttc atatctagac aaacccgggt gtcttgcatt tattagtact gtgagtttac
     3961 atacaaacga ggcttactca gataacatag agtcacctct aattagcaaa aacccatttc
     4021 taattagcta tttcgaccag ttctaaagtc accgtaatta gaaatggttt ttgtagcttt
     4081 tctaattaga aatgaccccg agataagtga aatatggcat tttctgggtt ttctagtaag
     4141 aaatgatccc gcacgtagtg attccgcgtt acaagccaaa ttcaaattaa aaaagtatat
     4201 tacacaattc aagttacatt attttcagga aacaatggcg aagagcaaag cagagattca
     4261 aaaggaggaa agagacaaca gatatgaacg aagacttctt gaaaagagaa agagacatga
     4321 ggtgagtaac catgataata aataatcaag taagggctat agatttataa attctggagt
     4381 taacagtttc gaaaaaaagt agttttcata tgtgcgaagg aatatcaaaa ataggggaat
     4441 agcctaaacg agtgagaaaa ttaaatgttt gttatttttt caaaaatgtc tgcagggccg
     4501 ttatcctttc aaatgcactc tatcaaacat agatcatatc catcatatct tatccagaat
     4561 ttatatcata tccaagattg agagcgtttc aaggaaaatc tcaaacaacc atacaatttt
     4621 gtagatcgac aacaaggcct ccacttcttc atcgatggcc tctgcagcac ctgcaccacc
     4681 aagtttgtat ttttcatatt aatataaagt gagccacgca tagtcgtttt taggattttt
     4741 gccagatcga tgtcgcgcaa acagactgtt tttttttctt tgaaagccgt gcgtgacatg
     4801 ggccacggat tgtaacgccg cgctggatgg ataaaaattt attcagtttt ttagccttta
     4861 caaaatatgg ttcgactact ttctttattt ttacattttt ataggtgctc attgagctga
     4921 gatattttgg gtataatcgg aaaatcgctg attttggtca tttttctgca ctaaagatta
     4981 cacaaaaagt tcagtattta gattcattgt tgaaaattgg ggatgatcta cagatacata
     5041 tcgccgtgtt caaaaacaca cgtgctggcg ttacccaact taatcgcctt gcagcacatc
     5101 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5161 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     5221 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5281 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5341 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5401 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5461 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5521 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5581 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5641 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5701 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5761 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5821 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5881 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5941 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6001 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6061 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6121 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6181 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6241 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6301 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6361 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6421 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6481 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6541 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6601 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6781 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6841 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7021 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7081 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7201 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7261 ttttacggtt cctggtcttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7321 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccagc ggataacaat
     7381 ttcaca
//
