LOCUS       sequence-370713-        9502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector for insertion into 5' miniMos Arm; carries floxed 
            Cbr-unc-119 marker, Phsp16.41::Cre and Pmyo-2::2xNLS::cyOFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9502)
  AUTHORS   Schwartz ML, Davis MW, Rich MS, Jorgensen EM
  TITLE     High-efficiency CRISPR gene editing in C. elegans using Cas9 
            integrated into the genome.
  JOURNAL   PLoS Genet. 2021 Nov 8;17(11):e1009755. doi: 
            10.1371/journal.pgen.1009755. eCollection 2021 Nov.
  PUBMED    34748534
REFERENCE   2  (bases 1 to 9502)
  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: "PLoS 
            Genet."; date: "2021-11-8"; volume: "17(11):e1009755. doi"; pages: "
            10.1371/journal.pgen.1009755. eCollection 2021 Nov"
COMMENT     Sequence Label: pMLS791
FEATURES             Location/Qualifiers
     source          1..9502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     201..218
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     202..218
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     228..247
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        228..246
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     271..287
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     272..288
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1902..2225
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2334..2354)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2334..2352)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2373..2389)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2373..2389)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3376..3396
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4290..4361
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    4916..4949
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             6909..6920
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    7069..7102
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     7573..7595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7633..7651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7719..7823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7824..7892,7893..8684)
                     /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(8042..8061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8855..9443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9344..9363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccagc ggataacaat ttcacatcag gtgtacaagt atgaaatgtc gtttcgaaca
      121 tatagatgtc tcgcaaacgt aaatatttat cgattgtcat aaaactttga ccttgtgaag
      181 tgtcaacctt gactgtcgaa tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
      241 tatagggcga attgggtacc gggccccccc tcgaggtcga cggtatcgat aagcttgatt
      301 acatcgccga ccttgaactc gaccgtcgcg ccgaccgtct ttaatttcag aaaattcggc
      361 gcgatcacca aaaacggaac gttgagctgg acggaaatag tggtaaagtg acatgattat
      421 agtttgaaga tttctaattt cacaattaga gcaaatgttg ttcggtattt attttcaacg
      481 gtatttatac tattttccac ctttttctag aacattcgag ctgcttgttg caaaaggagg
      541 gcgactcaca ttcggtacat ggaaaagtag tgtacacaat aaagagaccc agatacattt
      601 tccgtctgcg tctctttgca cccaccggga gtattttcaa acgaatgcat ctaggacctt
      661 ctagaacatt ctgtaaggct gcagaatgcg ggtatataag gaaagcgggc tcagaggaag
      721 ccaacacgct ttgttctagt gcatctaaaa aacttcgaaa atatgtcaaa tcttctcact
      781 gttcatcaaa atcttccagc acttccagtt gacgcgacaa gtgatgaagt tcgcaagaac
      841 ctcatggata tgttccgtga cagacaagcc ttctccgagc acacctggaa gatgcttctc
      901 tcagtgtgcc gttcttgggc tgcctggtgc aagcttaaca accgcaagtg gttcccagcc
      961 gagccagaag acgttcgtga ctacttgctc tacctccaag ctcgtggatt ggcggtgaag
     1021 acgatccaac aacatctcgg acagctcaat atgctccatc gccgatctgg acttccacgc
     1081 ccatcagatt ctaatgccgt atctctcgta atgagacgta tccgcaagga aaatgttgat
     1141 gctggagaga gagccaagca agccctcgcc ttcgaaaggt aagtttaaat attaatgtat
     1201 tcaatcctaa gacttgacga aattttcaga accgacttcg atcaggtccg ttctcttatg
     1261 gaaaattccg atcgttgcca ggacatccga aacctcgcct tcctcggaat tgcttacaac
     1321 acacttctcc gtatcgccga aattgcccgc attcgcgtca aggatattag tcgcacagac
     1381 ggaggaagaa tgctcatcca catcggacgt acaaaaacgc tggtctcgac tgccggagtc
     1441 gaaaaagcgc tctcccttgg agttaccaag ctcgttgagc gctggatttc ggtttccgga
     1501 gtcgccgatg atccaaacaa ctacctgttc tgtcgtgttc gcaagaacgg agttgctgcc
     1561 ccgagtgcaa catcacaact ctcgactcga gctcttgaag gtaagtttaa acaacatatt
     1621 agctaagaat aagctgaaaa taaattttca ggaatcttcg aagctaccca ccgtctcatc
     1681 tacggagcca aggatgattc cggacaacgt tacttggctt ggtctggaca ttccgcccgt
     1741 gtcggagctg cccgtgacat ggctcgcgct ggagtttcca tcccagagat tatgcaagcc
     1801 ggaggttgga ccaacgtcaa cattgtgatg aactatatta gaaacttgga ttctgaaact
     1861 ggcgccatgg ttcgtcttct cgaagacggt gattaagata aatgcaaaat cctttcaagc
     1921 attcccttct tctctatcac tcttctttct ttttgtcaaa aaattctctc gctaatttat
     1981 ttgctttttt aatgttatta ttttatgact ttttatagtc actgaaaagt ttgcatctga
     2041 gtgaagtgaa tgctatcaaa atgtgattct gtctgatgta ctttcacaat ctctcttcaa
     2101 ttctattttg aagtgcttta aacccgaaag gttgagaaaa atgcgagcgc tcaaatattt
     2161 gtattgtgtt cgttgagtga cccaacaaaa agaggaaact ttattgtgcc gccaagaaaa
     2221 aagtctcaca taaatcaata tctcacatat cttgccgatt cggcttgcgc ctgcagattc
     2281 tcttccgtcc aatcgaattg gccgccaccg cggtggagct ccagcttttg ttccctttag
     2341 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgc cattttatat
     2401 ctgagtagta tcctttgctt taaatgtcca taaaactaat tttataatca ataaaacaac
     2461 gtttgtaaat caactgagtt tacaagtaga gacattgagg gatactttca ctatgctaaa
     2521 gtgataatcg accaaataat aacttcactt tggtatttat tcctgtctta taaatgttat
     2581 gtatgaatta aattcatatg catatggctc actctgacaa tttttaataa tcttccagat
     2641 caatattgac taccgatgcg ggtggtcttt tgctttgaat tctgctgaac tttacacccc
     2701 gaacagcaat gtgtgcttca gcctaaaaaa aagtaagtgt gttaatcagt gcccccgatt
     2761 cttcattttt tgcccctctc tcccgtttcg tcggcaaaag aagagaaaat aaagataagt
     2821 ctcaagatag gttggtaatc gctaaagtgg ttgtgtggat aagagtagca aaatggcagg
     2881 aagagcactt tgcgcgcaca cactgtactc attgttctgg ataaaattct ctcgttgttt
     2941 gccgtcggat gtctgcctct ctgcattgag ccggcttctt cactatcttt agttaaccta
     3001 aaatgccgtt tcttttctcg tatccccact atcccgttga ggttctctgc tctcttcgct
     3061 ccctaccgcc agcgagcaac tatccgtggg ggcgccttgc tcggaagatg ggggggaaga
     3121 aagaagattt ttgctatttg cacttgagaa agagactttt cctgcgtcga tggtcagaga
     3181 acagtgtgca gacacttttc agctacctag aattacaatt ggatatcccc gcctcccaat
     3241 ccacccaccc agggaaaaag aagggctcgc cgaaaatcaa agttatctcc aggctcgcgc
     3301 atcccaccga gcggttgact tctctccacc acttttcatt ttaaccctcg atcgtcagac
     3361 acagaaatgg gatccccaaa gaagaagcgt aaggtgtcca aggtaacact tagtttttgt
     3421 tgctctgttt aaaaattaat tttaagggag aggagctcat caaggagaac atgcgttcca
     3481 agctctacct cgagggatcc gtcaacggac accaattcaa gtgcacccac gagggagagg
     3541 gaaagccata cgagggaaag caaaccaacc gtatcaaggt cgtcgaggga ggaccactcc
     3601 cattcgcctt cgacatcctc gccacccact tcatgtacgg atccaaggta agttttcttg
     3661 agacctttac taactaacta ggtctcatag attttcaggt cttcatcaag tacccagccg
     3721 acctcccaga ctacttcaag caatccttcc cagagggatt cacctgggag cgtgtcatgg
     3781 tcttcgagga cggaggagtc ctcaccgcca cccaagacac ctccctccaa gacggagagc
     3841 tcatctacaa cgtcaaggta agttatgtag agaccttaat actcaactat ggtctcaaac
     3901 attttcaggt ccgtggagtc aacttcccag ccaacggacc agtcatgcaa aagaagaccc
     3961 tcggatggga gccatccacc gagaccatgt acccagccga cggaggactc gagggacgtt
     4021 gcgacaaggc cctcaagctc gtcggaggag gacacctcca cgtcaacttc aaggtaagtt
     4081 ctttagagac ctaatcacat caatttggtc tcagtaattt tcagaccacc tacaagtcca
     4141 agaagccagt caagatgcca ggagtccact acgtcgaccg tcgtctcgag cgtatcaagg
     4201 aggccgacaa cgagacctac gtcgagcaat acgagcacgc cgtcgcccgt tactccaacc
     4261 tcggaggagg aatggacgag ctctacaagt cccgtcgtcg taaggccaac ccaaccaagc
     4321 tctccgagaa cgccaagaag ctcgccaagg aggtcgagaa ctaaggatga tcgacgccaa
     4381 cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta ttatttattt
     4441 attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa atctaaacat
     4501 ttttttttct ctttccgtct cccaattcgt attccgctcc tctcatctga acacaatgtg
     4561 caagtttatt tatcttctcg ctttcatttc attaggacgt ggggggaatt ggtggaaggg
     4621 ggaaacacac aaaaggatga tggaaatgaa ataaggacac acaatatgca acaacattca
     4681 attcagaaat atggaggaag gtttaaaaga aaacataaaa atatatagag gaggaaggaa
     4741 aactagtaaa aaataagcaa agaaattagg cgaacgatga gaattgtcct cgcttggcta
     4801 ggtgagctct ggtaccctct agtcaaggcc ttaagtgagt cgtattacgt agcccgggcc
     4861 tagttctagt ttaaaccacg tgcgtgagcg taagtttaaa catgatttta ctaacataac
     4921 ttcgtataaa gtatcctata cgaagttatg acattctcta atgaaaaaat ctttcagttg
     4981 aaattgaaaa tgagttaaag ttggagtttt tattgaaaac agatttccgt gtgattagtg
     5041 tttttagcga gtgtgacagg acagcgaaaa aatatagaaa caagggggga actgaaaagc
     5101 ttaggaatgc attgaacatg agaaggggaa agggaaggaa caaactagac aggaattatt
     5161 ggaatttaat cacatttgga gttttttttt ctattcgaca gaataattat ccagaacatt
     5221 tttgtattaa atatttatgc atcatatgag tagtcggctt tgttgtgcat gacgagtttg
     5281 ttatcgacga aatagaagct gtcagaacga gtctcgtttg gattgttgat catgtcgtcc
     5341 actgaaaaag agattagtct tttgaattgt actttttaga ataatgactc actgagttgt
     5401 tgagagagtt gagggaactc atagatatgt tcacagttgt ttcgtgaatt cggaatacag
     5461 aatccgaatt caaagtcaaa acacttcaga aggcgatctt tgaagaagtg acgttcgatc
     5521 attcggaaat gatggattgg gatgtctcct actttgaatt ccacagttgc tccgaccgtt
     5581 ttgagtttca aaaagtttgg agcaaatcta taacgcacgt atcttgccga ctcctgtggc
     5641 gactcatcat tctcctgatc gttctccggt ttggcgatct cgaaaagcac ttgctcagtg
     5701 tccagatcac ggatttggaa cttggtgaac tcgatgttat agatgttcgc agatggggag
     5761 cataagaatc ctaaatttat gttttaaact gaaatccaaa gggagcaaga taccttgagt
     5821 gattcccgga agtgctaaaa cgtcgttcgg agtgatttga gctttcttcg caagctccga
     5881 ttccgttgtg attccttgtt cggtgcttgg tggtggccgt ggcatctgga aatatggaaa
     5941 agttcaacaa aaagaaaaga gaaaagaatg aaatcggata tcaagagtta gttgagcggt
     6001 ttctctagtt ttctgagtct cacctgcgac gggaaggtcg ccgagccggg tggaatcgat
     6061 cgttgttgct cggctttcat atcggtttgg ttggaagcgg ctgaaaacgg aaagaagtgg
     6121 aagaaggaaa agagtgtggt gtgacaggaa aatggtaatt agagggtgcc aaataaccag
     6181 ctatattttg tttttttttg aaaacatttt taaaaagaaa aatacgataa tgatatcaga
     6241 tggatttccg gaaaactggt atgaaaaatt tcaacctttt tgagtacatg taatcaaaat
     6301 acactttgta aattatcatt tttattgaaa ctccaccatt tttctattta taacgctaat
     6361 aatttgaaaa agaaacctat tgcgaaccgc ggggtgaatc ccaaaaacga atgcgttttg
     6421 gtggagtgat tgattcgaat cgaagaagaa aaagaagaag acgtggaata gagagctcac
     6481 tcttaaccga gcagcacaca ccgacagaaa aaaaaatgaa atgaatgagg gtcttcttct
     6541 tcttcttctt cgaatgattg acagaaatgg gaaaaagagg aagattgaga agggaaaaag
     6601 gaaggagaaa agaagcagaa gaagacgtca gagaggagag gaacgagcgg aaaagcagcg
     6661 ggcgcaagtc atagaagtag cagagctggg gagaagaaga cactatccaa gaaaggaatg
     6721 acgagagagt atgcaaaggg gtatagggtg cagacagaat aggaacagaa taacagatga
     6781 tgagccaaga agagttgaaa agggcgatga atttgtcatg taacttaatt tgggtcaatt
     6841 tgagcatgat gaattgaaat catcccttgt tgggagttaa taaccggttt gttatcagaa
     6901 accctgtaat agaagggcgc cctaactttg agccaattca tcccggtttc tgtcaaatat
     6961 atcaaaaagt ggtcaactga caaattgttt ttgatattat aataaacatt ttatccgtta
     7021 acaattttcg aatacttttt acaaggactt ggataaattg gctcaaagat aacttcgtat
     7081 aaagtatcct atacgaagtt attaactaat ctgatttaaa ttttcgcgaa ttgagcgtca
     7141 taattgttta ctctcagtgc agtcaacatg tcgagtttcg tgccgaataa agagcaaacg
     7201 cggacagtat taattttctg ttttcatttg aagaaaacag ctgcggacgt cgtgactggg
     7261 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     7321 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     7381 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     7441 ggtgcactct cagtgcaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     7501 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     7561 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     7621 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     7681 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     7741 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     7801 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     7861 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     7921 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     7981 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     8041 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     8101 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     8161 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     8221 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     8281 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     8341 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     8401 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     8461 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     8521 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggtca gatggtaagc
     8581 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     8641 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     8701 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     8761 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8821 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8881 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8941 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     9001 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     9061 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     9121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     9181 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     9241 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     9301 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     9361 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     9421 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggtct
     9481 tttgctggcc ttttgctcac at
//
