LOCUS       sequence-370784-        7095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector for insertion into 5' miniMos Arm; carries floxed 
            Cbr-unc-119 marker and Phsp16.41::Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7095)
  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 7095)
  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: pMLS716
FEATURES             Location/Qualifiers
     source          1..7095
                     /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"
     protein_bind    2509..2542
                     /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             4502..4513
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    4662..4695
                     /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     5166..5188
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5226..5244)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5312..5416
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5417..5485,5486..6277)
                     /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(5635..5654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6448..7036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6937..6956
                     /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 ctaggtgagc
     2401 tctggtaccc tctagtcaag gccttaagtg agtcgtatta cgtagcccgg gcctagttct
     2461 agtttaaacc acgtgcgtga gcgtaagttt aaacatgatt ttactaacat aacttcgtat
     2521 aaagtatcct atacgaagtt atgacattct ctaatgaaaa aatctttcag ttgaaattga
     2581 aaatgagtta aagttggagt ttttattgaa aacagatttc cgtgtgatta gtgtttttag
     2641 cgagtgtgac aggacagcga aaaaatatag aaacaagggg ggaactgaaa agcttaggaa
     2701 tgcattgaac atgagaaggg gaaagggaag gaacaaacta gacaggaatt attggaattt
     2761 aatcacattt ggagtttttt tttctattcg acagaataat tatccagaac atttttgtat
     2821 taaatattta tgcatcatat gagtagtcgg ctttgttgtg catgacgagt ttgttatcga
     2881 cgaaatagaa gctgtcagaa cgagtctcgt ttggattgtt gatcatgtcg tccactgaaa
     2941 aagagattag tcttttgaat tgtacttttt agaataatga ctcactgagt tgttgagaga
     3001 gttgagggaa ctcatagata tgttcacagt tgtttcgtga attcggaata cagaatccga
     3061 attcaaagtc aaaacacttc agaaggcgat ctttgaagaa gtgacgttcg atcattcgga
     3121 aatgatggat tgggatgtct cctactttga attccacagt tgctccgacc gttttgagtt
     3181 tcaaaaagtt tggagcaaat ctataacgca cgtatcttgc cgactcctgt ggcgactcat
     3241 cattctcctg atcgttctcc ggtttggcga tctcgaaaag cacttgctca gtgtccagat
     3301 cacggatttg gaacttggtg aactcgatgt tatagatgtt cgcagatggg gagcataaga
     3361 atcctaaatt tatgttttaa actgaaatcc aaagggagca agataccttg agtgattccc
     3421 ggaagtgcta aaacgtcgtt cggagtgatt tgagctttct tcgcaagctc cgattccgtt
     3481 gtgattcctt gttcggtgct tggtggtggc cgtggcatct ggaaatatgg aaaagttcaa
     3541 caaaaagaaa agagaaaaga atgaaatcgg atatcaagag ttagttgagc ggtttctcta
     3601 gttttctgag tctcacctgc gacgggaagg tcgccgagcc gggtggaatc gatcgttgtt
     3661 gctcggcttt catatcggtt tggttggaag cggctgaaaa cggaaagaag tggaagaagg
     3721 aaaagagtgt ggtgtgacag gaaaatggta attagagggt gccaaataac cagctatatt
     3781 ttgttttttt ttgaaaacat ttttaaaaag aaaaatacga taatgatatc agatggattt
     3841 ccggaaaact ggtatgaaaa atttcaacct ttttgagtac atgtaatcaa aatacacttt
     3901 gtaaattatc atttttattg aaactccacc atttttctat ttataacgct aataatttga
     3961 aaaagaaacc tattgcgaac cgcggggtga atcccaaaaa cgaatgcgtt ttggtggagt
     4021 gattgattcg aatcgaagaa gaaaaagaag aagacgtgga atagagagct cactcttaac
     4081 cgagcagcac acaccgacag aaaaaaaaat gaaatgaatg agggtcttct tcttcttctt
     4141 cttcgaatga ttgacagaaa tgggaaaaag aggaagattg agaagggaaa aaggaaggag
     4201 aaaagaagca gaagaagacg tcagagagga gaggaacgag cggaaaagca gcgggcgcaa
     4261 gtcatagaag tagcagagct ggggagaaga agacactatc caagaaagga atgacgagag
     4321 agtatgcaaa ggggtatagg gtgcagacag aataggaaca gaataacaga tgatgagcca
     4381 agaagagttg aaaagggcga tgaatttgtc atgtaactta atttgggtca atttgagcat
     4441 gatgaattga aatcatccct tgttgggagt taataaccgg tttgttatca gaaaccctgt
     4501 aatagaaggg cgccctaact ttgagccaat tcatcccggt ttctgtcaaa tatatcaaaa
     4561 agtggtcaac tgacaaattg tttttgatat tataataaac attttatccg ttaacaattt
     4621 tcgaatactt tttacaagga cttggataaa ttggctcaaa gataacttcg tataaagtat
     4681 cctatacgaa gttattaact aatctgattt aaattttcgc gaattgagcg tcataattgt
     4741 ttactctcag tgcagtcaac atgtcgagtt tcgtgccgaa taaagagcaa acgcggacag
     4801 tattaatttt ctgttttcat ttgaagaaaa cagctgcgga cgtcgtgact gggaaaaccc
     4861 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4921 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     4981 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     5041 tctcagtgca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     5101 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5161 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     5221 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5281 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5341 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5401 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5461 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5521 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5581 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5641 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5701 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5761 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5821 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5881 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5941 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6001 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6061 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6121 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg tcagatggta agccctcccg
     6181 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6241 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6301 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6361 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6421 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6481 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6541 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6601 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6661 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6721 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6781 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6841 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6901 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6961 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7021 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg tcttttgctg
     7081 gccttttgct cacat
//
