LOCUS       sequence-382980-        7476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a plasmid modified from miniMOS vector pCFJ910. the modification 
            including insertions of myo2p::TagRFP downstream to NeoR CDS and 2 
            loxP sites flanking the NeoR and tagRFP cassettes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7476)
  AUTHORS   Li J, Qin Y, Shen C, Zhang J, Tu S, Yang J, Wang Y, Zhou R, Zhang K,
            Chen J, Yang W
  TITLE     A new miniMOS tool kit capable of visualizing single copy insertion 
            in C. elegans.
  JOURNAL   PeerJ. 2023 May 17;11:e15433. doi: 10.7717/peerj.15433. eCollection 
            2023.
  PUBMED    37214099
REFERENCE   2  (bases 1 to 7476)
  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: "PeerJ."; 
            date: "2023-05-17"; volume: "11:e15433. doi"; pages: " 
            10.7717/peerj.15433. eCollection 2023"
COMMENT     Sequence Label: YW347_Ptbb2UTR-loxP-RFP-miniMOS
FEATURES             Location/Qualifiers
     source          1..7476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     283..299
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     283..299
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      341..664
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(695..728)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1566..2360
                     /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(1620..1639)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2230..2249
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             3339..4070
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQDIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     protein_bind    complement(4778..4811)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5480..5502
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5540..5558)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5626..5730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5731..5799,5800..6591)
                     /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(5949..5968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6762..7350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7251..7270
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaccgtacg tctcgaggaa ttctatcagg tgtacaagta tgaaatgtcg tttcgaacat
       61 atagatgtct cgcaaacgta aatatttatc gattgtcata aaactttgac cttgtgaagt
      121 gtcaaccttg actgtcgaac caccatagtt tggcgcgaat tgagcgtcat aattgtttac
      181 tctcagtgca gtcaacatgt cgagtttcgt gccgaataaa gagcaaacgc ggacagtatt
      241 aattttctgt tttcatttga agaaaacagc tgcggccgga tccaggaaac agctatgacc
      301 atgattacgc caagctacgt aatacgactc actaggataa atgcaaaatc ctttcaagca
      361 ttcccttctt ctctatcact cttctttctt tttgtcaaaa aattctctcg ctaatttatt
      421 tgctttttta atgttattat tttatgactt tttatagtca ctgaaaagtt tgcatctgag
      481 tgaagtgaat gctatcaaaa tgtgattctg tctgatgtac tttcacaatc tctcttcaat
      541 tccattttga agtgctttaa acccgaaagg ttgagaaaaa tgcgagcgct caaatatttg
      601 tattgtgttc gttgagtgac ccaacaaaaa gaggaaactt tattgtgccg ccaagaaaaa
      661 agtcctcgag gtcgacaggc ctgcatgcgt cgacataact tcgtatagca tacattatac
      721 gaagttatag ttgcaggacc actgcggccg catgggccat ggcaccttaa catttctaaa
      781 ttttagttta ataaaaaaat tttaaaataa tacgagtttt gagactaact ttcaataaaa
      841 accaaaaaat tgcttcaaaa aagagctcgc gccgatttac gataaacaca gcgcgctacg
      901 acactccgaa aaccgcgccg caccattaca ttctccgccc caccgcacgg tttgcggaag
      961 gcgatgaaat tgctgggcct aaaattttaa cccttttttt cccgtttttc aactgttttc
     1021 ccgatttttt cgatattttt ttcaggtttt tagttagaaa agtatataaa aattgcatta
     1081 tttgtagttt tcgctcgtaa tttgtgatag aaaacggcca atttgcctag atttgtgatt
     1141 ctaccaagtg gaatttttgg gaaaatatca atattccacg atttttattc aaatcataac
     1201 gtttcgaaat tatcagctta aataaaacta aaataaataa taaaatacta aaaaccctga
     1261 atttctcggt tctcggttca gcttgagttc gttctataca cgttatcgat ttttgtcatt
     1321 tcgtttaatc atgggttttg attcatttta aatatgaaat ttgatttatt cctcgtttcg
     1381 aagttggttt gtcttgtccg gccggtaaac aaatgttatt aaaaaaaata ataaatttcg
     1441 aaatttttcg taatgttccg caatcgtgtt tttccgttta tctcgtcttc ccttccttca
     1501 aatatactat ttattttctc aattcacata ttttcagcca gccctgctca acaagtggaa
     1561 taaaaatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     1621 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     1681 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     1741 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     1801 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     1861 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     1921 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     1981 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     2041 acgaagagca tcaggggctc gcaccagccg aactgttcgc caggctcaag gcgcgcatgc
     2101 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     2161 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     2221 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     2281 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     2341 ttcttgacga gttcttctag cattttatat ctgagtagta cctttgcttt aaatgtccat
     2401 aaaactaatt ttataatcaa taaaacaacg tttgtaaatc aactgagttt acaagtagag
     2461 acattgaggg atactttcac tatgctaaag tgataatcga ccaaataata acttcacttt
     2521 ggtatttatt cctgtcttat aaatgttatg tatgaattaa attcatatgc atatggctca
     2581 ctctgacaat ttttaataat cttccagatc aatattgact accgatgcgg gtggtctttt
     2641 gctttgaatt ctgctgaact ttacaccccg aacagcaatg tgtgcttcag cctaaaaaaa
     2701 agtaagtgtg ttaatcagtg cccccgattc ttcatttttt gcccctctct cccgtttcgt
     2761 cggcaaaaga agagaaaata aagataagtc tcaagatagg ttggtaatcg ctaaagtggt
     2821 tgtgtggata agagtagcaa aatggcagga agagcacttt gcgcgcacac actgtactca
     2881 ttgttctgga taaaattctc tcgttatttg ccgtcggatg tctgcctctc tgcattgagc
     2941 cggcttcttc actatcttta gttaacctaa aatgccgttt cttttctcgt atccccacta
     3001 tcccgttgag gttctctgct ctcttcgctc cctaccgcca gcgagcaact atccgtgggg
     3061 gcgccttgct cggaagatgg gggggaagaa agaagatttt tgctatttgc acttgagaaa
     3121 gagacttttc ctgcgtcgat ggtcagagaa cagtgtgcag acacttttca gctacctaga
     3181 attacaattg gatatccccg cctcccaatc cacccaccca gggaaaaaga agggctcgcc
     3241 gaaaatcaaa gttatctcca ggctcgcgca tcccaccgag cggttgactt ctctccacca
     3301 cttttcattt taaccctcga tcgtcagaca cagaaatgat ggtgtctaag ggcgaagagc
     3361 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
     3421 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
     3481 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt
     3541 acggcagcag aaccttcatc aaccacaccc aggacatccc cgatttcttt aagcagtcct
     3601 tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg
     3661 ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
     3721 tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccaaca
     3781 ccgagatgct gtaccccgct gacggcggcc tggaaggcag aaccgacatg gccctgaagc
     3841 tcgtgggcgg gggccacctg atctgcaact tcaagaccac atacagatcc aagaaacccg
     3901 ctaagaacct caagatgccc ggcgtctact atgtggacca cagactggaa agaatcaagg
     3961 aggccgacaa agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc
     4021 tccctagcaa actggggcac aaacttaatg gcatggacga gctgtacaag taagtccaat
     4081 tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat
     4141 tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc ttttaagcca cctctaacaa
     4201 tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg
     4261 ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact
     4321 tcttatgttt tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa
     4381 aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc
     4441 tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa
     4501 tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc
     4561 ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga
     4621 tgagcacgat gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag
     4681 tagaagttga taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga
     4741 atacattccc aatataccaa acataactgt ttctcgaata acttcgtata gcatacatta
     4801 tacgaagtta tggtcgacgg tatcgataag cttgatccgg atctgcggct tactcaccag
     4861 acctggcccc atccgattac cacctattcg cttcgatggg acacgcactc gctgagcagc
     4921 gcttcgattc ttacgaaagt gtgaaaaaat ggctcgatga atggttcgcc gcaaaagacg
     4981 atgagttcta ctggcgtgga atccacaaat tgcccgagag atgggaaaaa tgtgtagcta
     5041 gcgacggcaa atactttgaa taaatgattt tttctttttc cacaaaattt aacgtgtttt
     5101 ttgatttaaa aaaaacgaca tttcatactt gtacacctga tatctagata cctaggtgag
     5161 ctctggtacc cccctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     5221 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5281 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5341 cgcatatggt gcactctcag tgcaatctgc tctgatgccg catagttaag ccagccccga
     5401 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5461 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5521 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5581 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5641 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5701 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5761 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5821 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5881 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5941 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6001 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6061 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6121 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6181 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6241 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6301 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6361 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6421 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggtcagat
     6481 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6541 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6601 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6661 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6721 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6841 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6901 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7141 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7261 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7321 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7381 ctggtctttt gctggccttt tgttcacatg ttctttcctg cgttatcccc tgattctgtg
     7441 gataaccgta ttaccgcctt tgagtgagct gatacc
//
