LOCUS       sequence-258304-        7588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  modified pCFJ151 for C. elegans mosSCI at ttTi5605 or oxTi354 in 
            otherwise wild-type background; floxed NeoR selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7588)
  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 7588)
  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: pALS297 (ttTi5605-MCS
            loxP::Prps-27::NeoR::unc...
FEATURES             Location/Qualifiers
     source          1..7588
                     /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             2249..3043
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2303..2322)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2913..2932
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(3746..3779)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5573..5595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5633..5651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5719..5823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5824..5892,5893..6684)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6042..6061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6855..7443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7344..7363
                     /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 tgcgcgtata acttcgtata gcatacatta tacgaagtta
     1441 tatggcacct taacatttct aaattttagt ttaataaaaa aattttaaaa taatacgagt
     1501 tttgagacta actttcaata aaaaccaaaa aattgcttca aaaaagagct cgcgccgatt
     1561 tacgataaac acagcgcgct acgacactcc gaaaaccgcg ccgcaccatt acattctccg
     1621 ccccaccgca cggtttgcgg aaggcgatga aattgctggg cctaaaattt taaccctttt
     1681 tttcccgttt ttcaactgtt ttcccgattt tttcgatatt tttttcaggt ttttagttag
     1741 aaaagtatat aaaaattgca ttatttgtag ttttcgctcg taatttgtga tagaaaacgg
     1801 ccaatttgcc tagatttgtg attctaccaa gtggaatttt tgggaaaata tcaatattcc
     1861 acgattttta ttcaaatcat aacgtttcga aattatcagc ttaaataaaa ctaaaataaa
     1921 taataaaata ctaaaaaccc tgaatttctc ggttctcggt tcagcttgag ttcgttctat
     1981 acacgttatc gatttttgtc atttcgttta atcatgggtt ttgattcatt ttaaatatga
     2041 aatttgattt attcctcgtt tcgaagttgg tttgtcttgt ccggccggta aacaaatgtt
     2101 attaaaaaaa ataataaatt tcgaaatttt tcgtaatgtt ccgcaatcgt gtttttccgt
     2161 ttatctcgtc ttcccttcct tcaaatatac tatttatttt ctcaattcac atattttcag
     2221 ccagccctgc tcaacaagtg gaataaaaat gattgaacaa gatggattgc acgcaggttc
     2281 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2341 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2401 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     2461 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     2521 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     2581 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     2641 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     2701 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcaccag ccgaactgtt
     2761 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     2821 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     2881 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     2941 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3001 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagtccaat tactcttcaa
     3061 catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat tatcaaaaaa
     3121 cttctcttaa tttctttgtt ttttagcttc ttttaagcca cctctaacaa tgaaattgtg
     3181 tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc
     3241 ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact tcttatgttt
     3301 tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa aataccttat
     3361 catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc
     3421 atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa tttttcaatc
     3481 aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc ccctattgtt
     3541 tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat
     3601 gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga
     3661 taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga atacattccc
     3721 aatataccaa acataactgt ttctcataac ttcgtatagc atacattata cgaagttata
     3781 gatctgccca ctagtgagtc gtattataag tgcaagtaag atcagtgttt gtttcgatat
     3841 cagtctgttt cgtaacggtc ttctgtataa ctacaaaaaa gaataaaaaa ccgtatcaca
     3901 actgagaaaa aagacaaaac ttattgaaca gtgttccttt caaaaaatta ttttgtaaaa
     3961 aatcgagttt ttctccaaaa taacgcacgc aatcaaaaac aagaaaacta acgtgttctc
     4021 ctgggcgtgg aactccttat caagattacc attattgtct gtcagaattg atatttcaat
     4081 tatttcatat ctagacaaac ccgggtgtct tgcatttatt agtactgtga gtttacatac
     4141 aaacgaggct tactcagata acatagagtc acctctaatt agcaaaaacc catttctaat
     4201 tagctatttc gaccagttct aaagtcaccg taattagaaa tggtttttgt agcttttcta
     4261 attagaaatg accccgagat aagtgaaata tggcattttc tgggttttct agtaagaaat
     4321 gatcccgcac gtagtgattc cgcgttacaa gccaaattca aattaaaaaa gtatattaca
     4381 caattcaagt tacattattt tcaggaaaca atggcgaaga gcaaagcaga gattcaaaag
     4441 gaggaaagag acaacagata tgaacgaaga cttcttgaaa agagaaagag acatgaggtg
     4501 agtaaccatg ataataaata atcaagtaag ggctatagat ttataaattc tggagttaac
     4561 agtttcgaaa aaaagtagtt ttcatatgtg cgaaggaata tcaaaaatag gggaatagcc
     4621 taaacgagtg agaaaattaa atgtttgtta ttttttcaaa aatgtctgca gggccgttat
     4681 cctttcaaat gcactctatc aaacatagat catatccatc atatcttatc cagaatttat
     4741 atcatatcca agattgagag cgtttcaagg aaaatctcaa acaaccatac aattttgtag
     4801 atcgacaaca aggcctccac ttcttcatcg atggcctctg cagcacctgc accaccaagt
     4861 ttgtattttt catattaata taaagtgagc cacgcatagt cgtttttagg atttttgcca
     4921 gatcgatgtc gcgcaaacag actgtttttt tttctttgaa agccgtgcgt gacatgggcc
     4981 acggattgta acgccgcgct ggatggataa aaatttattc agttttttag cctttacaaa
     5041 atatggttcg actactttct ttatttttac atttttatag gtgctcattg agctgagata
     5101 ttttgggtat aatcggaaaa tcgctgattt tggtcatttt tctgcactaa agattacaca
     5161 aaaagttcag tatttagatt cattgttgaa aattggggat gatctacaga tacatatcgc
     5221 cgtgttcaaa aacacacgtg ctgtgacgtc gtgactgggg aaaaccctgg cgttacccaa
     5281 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5341 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     5401 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtgcaatc
     5461 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5521 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5581 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5641 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5821 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5881 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5941 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6001 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6061 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6121 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6181 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6241 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6301 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6361 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6421 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6481 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6541 tctcgcggta tcattgcagc actggggtca gatggtaagc cctcccgtat cgtagttatc
     6601 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6661 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6721 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6781 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6841 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6901 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6961 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7021 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7081 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7141 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7201 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7261 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7321 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7381 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7441 aaaaacgcca gcaacgcggc ctttttacgg ttcctggtct tttgctggcc ttttgctcac
     7501 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7561 gctgatacca gcggataaca atttcaca
//
