LOCUS       sequence-370711-        9278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ttTi5605 MosSci targeting vector with floxed Cbr-unc-119 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9278)
  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 9278)
  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: pMLS621
FEATURES             Location/Qualifiers
     source          1..9278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     339..361
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(399..417)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        485..589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(590..658,659..1450)
                     /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(808..827)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1621..2209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2110..2129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3726..3759
                     /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             5718..5729
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    5878..5911
                     /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)."
     protein_bind    6065..6188
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6229..6534)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(6331..6350)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(6854..7534)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     7449..7468
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7588..7594,7595..7612,7613..7618))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(7643..7766)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7774..7791)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7774..7790)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cacgtgctgt gacgtcgtga ctggggaaaa ccctggcgtt acccaactta atcgccttgc
       61 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
      121 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
      181 tctgtgcggt atttcacacc gcatatggtg cactctcagt gcaatctgct ctgatgccgc
      241 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      301 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
      361 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
      421 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
      481 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      541 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      601 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      661 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      721 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      781 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      841 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      901 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      961 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1021 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1081 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1141 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1201 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1261 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1321 tgcagcactg gggtcagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1381 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1441 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1501 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1561 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1621 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1681 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1741 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1801 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1861 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1921 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1981 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2041 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2101 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2161 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2221 cgcggccttt ttacggttcc tggtcttttg ctggcctttt gctcacatgt tctttcctgc
     2281 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccagcgg
     2341 ataacaattt cacagggccc tgtcggatta tgggctcttc ttttatcctc tactttcttt
     2401 ctagaaaatg caaatgttga gtcagaaatt gctgaatgga gtacaactgc aatgggttta
     2461 gcttatgttt ctggaacaat tactggtgga attgtgattg aaaagtaagg ttatatttca
     2521 tttcaataaa tgttaccaca gtataacaaa aacaagttac attgttcaga gtatgtctga
     2581 atgtattatt attattgtat ttgtgcatca gtactgtagt tgtactgaag aagggtttaa
     2641 aataatctca acgtttttaa gacgagttag tgagaatacg gtcgcagtta ggatgtttgg
     2701 acatagtttt attagataat cgtgaactaa ctcactataa aaacccgcta actcattgaa
     2761 taagatagtt ttaaacgttc aaaatttgaa gacaaaattg tatgaataag tttacagagt
     2821 tggtcgacgc aagcttctcc ttctcttcac atttctcaat aatttagctc taattgcatt
     2881 cgtattcttc gcaaaaatac gtattctgat tgacccgatg aaatacggat gcctagttgc
     2941 attgatcatc tacggttata cttatgggtg aggtttttca aatatatttt gatttaatcg
     3001 aataatgaaa tttcagaact ggagttggtc caatttcgtg gtttatatcc tctgaactgg
     3061 tacctcaaaa acatcgtagt attgctcaat ctgtggcata ttcaatgaat actgtaatgg
     3121 tggttatcag tacatttaca gtattacctc tatattccgt tattggtagt tatgctttcc
     3181 ttattctata ttcaattcct tcatttataa gtatgcttat attatttcgc tacttaccgg
     3241 aaaccaaagg acgagagatt catgatattg tgaatgaact taaaaaaaaa tgaaaacctt
     3301 tgaaaacgtt catacggccg aaattttact tgatcgagcc aattgaactt ttgttaagac
     3361 ttacagaaaa gtgagagatt ttgttacaaa aaagtgagaa aaaagcgaaa atgaacagga
     3421 aaaagaagaa aaatacaccc gaagtatcat ataaaaatta aaaagattgt ttgacctggc
     3481 ggaacttttt tgcaaaaact ctgaatatcc atggcacatg cttcgtgcaa aacaggatcc
     3541 aaatgaatat caacaagaga ttcttgaaga cgacgagcca cttgctcaga gcattgctta
     3601 tctcgaatga gacccttgtt aaaatcattt ttcaaacatt atatgacatt gtcgaaatgt
     3661 cctcctgatt ccatgatggt agcaaactca cttcgtagcg taagtttaaa catgatttta
     3721 ctaacataac ttcgtataaa gtatcctata cgaagttatg acattctcta atgaaaaaat
     3781 ctttcagttg aaattgaaaa tgagttaaag ttggagtttt tattgaaaac agatttccgt
     3841 gtgattagtg tttttagcga gtgtgacagg acagcgaaaa aatatagaaa caagggggga
     3901 actgaaaagc ttaggaatgc attgaacatg agaaggggaa agggaaggaa caaactagac
     3961 aggaattatt ggaatttaat cacatttgga gttttttttc tattcgacag aataattatc
     4021 cagaacattt ttgtattaaa tatttatgca tcatatgagt agtcggcttt gttgtgcatg
     4081 acgagtttgt tatcgacgaa atagaagctg tcagaacgag tctcgtttgg attgttgatc
     4141 atgtcgtcca ctgaaaaaga gattagtctt ttgaattgta ctttttagaa taatgactca
     4201 ctgagttgtt gagagagttg agggaactca tagatatgtt cacagttgtt tcgtgaattc
     4261 ggaatacaga atccgaattc aaagtcaaaa cacttcagaa ggcgatcttt gaagaagtga
     4321 cgttcgatca ttcggaaatg atggattggg atgtctccta ctttgaattc cacagttgct
     4381 ccgaccgttt tgagtttcaa aaagtttgga gcaaatctat aacgcacgta tcttgccgac
     4441 tcctgtggcg actcatcatt ctcctgatcg ttctccggtt tggcgatctc gaaaagcact
     4501 tgctcagtgt ccagatcacg gatttggaac ttggtgaact cgatgttata gatgttcgca
     4561 gatggggagc ataagaatcc taaatttatg ttttaaactg aaatccaaag ggagcaagat
     4621 accttgagtg attcccggaa gtgctaaaac gtcgttcgga gtgatttgag ctttcttcgc
     4681 aagctccgat tccgttgtga ttccttgttc ggtgcttggt ggtggccgtg gcatctggaa
     4741 atatggaaaa gttcaacaaa aagaaaagag aaaagaatga aatcggatat caagagttag
     4801 ttgagcggtt tctctagttt tctgagtctc acctgcgacg ggaaggtcgc cgagccgggt
     4861 ggaatcgatc gttgttgctc ggctttcata tcggtttggt tggaagcggc tgaaaacgga
     4921 aagaagtgga agaaggaaaa gagtgtggtg tgacaggaaa atggtaatta gagggtgcca
     4981 aataaccagc tatattttgt ttttttttga aaacattttt aaaaagaaaa atacgataat
     5041 gatatcagat ggatttccgg aaaactggta tgaaaaattt caaccttttt gagtacatgt
     5101 aatcaaaata cactttgtaa attatcattt ttattgaaac tccaccattt ttctatttat
     5161 aacgctaata atttgaaaaa gaaacctatt gcgaaccgcg gggtgaatcc caaaaacgaa
     5221 tgcgttttgg tggagtgatt gattcgaatc gaagaagaaa aagaagaaga cgtggaatag
     5281 agagctcact cttaaccgag cagcacacac cgacagaaaa aaaaatgaaa tgaatgaggg
     5341 tcttcttctt cttcttcttc gaatgattga cagaaatggg aaaaagagga agattgagaa
     5401 gggaaaaagg aaggagaaaa gaagcagaag aagacgtcag agaggagagg aacgagcgga
     5461 aaagcagcgg gcgcaagtca tagaagtagc agagctgggg agaagaagac actatccaag
     5521 aaaggaatga cgagagagta tgcaaagggg tatagggtgc agacagaata ggaacagaat
     5581 aacagatgat gagccaagaa gagttgaaaa gggcgatgaa tttgtcatgt aacttaattt
     5641 gggtcaattt gagcatgatg aattgaaatc atcccttgtt gggagttaat aaccggtttg
     5701 ttatcagaaa ccctgtaata gaagggcgcc ctaactttga gccaattcat cccggtttct
     5761 gtcaaatata tcaaaaagtg gtcaactgac aaattgtttt tgatattata ataaacattt
     5821 tatccgttaa caattttcga atacttttta caaggacttg gataaattgg ctcaaagata
     5881 acttcgtata aagtatccta tacgaagtta ttaactaatc tgatttaaat tttcaggctc
     5941 acgcacgtgg tttaaactag aactaggccc gggctacgta atacgactca cttaaggcct
     6001 tgactagagg gtaccagagc tcacctaggc aggaacagct atgaccatga ttacgccaag
     6061 ctatcaactt tgtatagaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata
     6121 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
     6181 tcactatggt cgacctgcag actggctgtg tataagggag cctgacattt atattcccca
     6241 gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc
     6301 ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc
     6361 atccccgata tgcaccaccg ggtaaagttc acgggggact ttatctgaca gcagacgtgc
     6421 actggccagg gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc
     6481 cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc
     6541 agcccctgtt ctcgtcggca aaagagccgt tcatttcaat aaaccgggcg acctcagcca
     6601 tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca
     6661 tggttgtgct taccgaaccg gagatattga catcatatat gccttgagca actgatagct
     6721 gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt
     6781 cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac
     6841 tgttatctgg cttttagtaa gccggatcct ctagattacg ccccgcctgc cactcatcgc
     6901 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     6961 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     7021 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     7081 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     7141 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     7201 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     7261 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt
     7321 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     7381 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     7441 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     7501 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     7561 tcgacggatc ctaactcaaa atccacacat tatacgagcc ggaagcataa agtgtaaagc
     7621 ctggggtgcc taatgcggcc gccatagtga ctggatatgt tgtgttttac agtattatgt
     7681 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc
     7741 tcgttcaact ttattataca tagttgataa ttcactggcc gtcgttttac actcgagacg
     7801 tacggtgcgc gcgatgcatt cgaagatctg cccactagtg agtcgtatta taagtgcaag
     7861 taagatcagt gtttgtttcg atatcagtct gtttcgtaac ggtcttctgt ataactacaa
     7921 aaaagaataa aaaaccgtat cacaactgag aaaaaagaca aaacttattg aacagtgttc
     7981 ctttcaaaaa attattttgt aaaaaatcga gtttttctcc aaaataacgc acgcaatcaa
     8041 aaacaagaaa actaacgtgt tctcctgggc gtggaactcc ttatcaagat taccattatt
     8101 gtctgtcaga attgatattt caattatttc atatctagac aaacccgggt gtcttgcatt
     8161 tattagtact gtgagtttac atacaaacga ggcttactca gataacatag agtcacctct
     8221 aattagcaaa aacccatttc taattagcta tttcgaccag ttctaaagtc accgtaatta
     8281 gaaatggttt ttgtagcttt tctaattaga aatgaccccg agataagtga aatatggcat
     8341 tttctgggtt ttctagtaag aaatgatccc gcacgtagtg attccgcgtt acaagccaaa
     8401 ttcaaattaa aaaagtatat tacacaattc aagttacatt attttcagga aacaatggcg
     8461 aagagcaaag cagagattca aaaggaggaa agagacaaca gatatgaacg aagacttctt
     8521 gaaaagagaa agagacatga ggtgagtaac catgataata aataatcaag taagggctat
     8581 agatttataa attctggagt taacagtttc gaaaaaaagt agttttcata tgtgcgaagg
     8641 aatatcaaaa ataggggaat agcctaaacg agtgagaaaa ttaaatgttt gttatttttt
     8701 caaaaatgtc tgcagggccg ttatcctttc aaatgcactc tatcaaacat agatcatatc
     8761 catcatatct tatccagaat ttatatcata tccaagattg agagcgtttc aaggaaaatc
     8821 tcaaacaacc atacaatttt gtagatcgac aacaaggcct ccacttcttc atcgatggcc
     8881 tctgcagcac ctgcaccacc aagtttgtat ttttcatatt aatataaagt gagccacgca
     8941 tagtcgtttt taggattttt gccagatcga tgtcgcgcaa acagactgtt tttttttctt
     9001 tgaaagccgt gcgtgacatg ggccacggat tgtaacgccg cgctggatgg ataaaaattt
     9061 attcagtttt ttagccttta caaaatatgg ttcgactact ttctttattt ttacattttt
     9121 ataggtgctc attgagctga gatattttgg gtataatcgg aaaatcgctg attttggtca
     9181 tttttctgca ctaaagatta cacaaaaagt tcagtattta gattcattgt tgaaaattgg
     9241 ggatgatcta cagatacata tcgccgtgtt caaaaaca
//
