LOCUS       sequence-213754-        8495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cloning expression cassettes by HR in yeast, Universal MosSCI 
            compatible.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8495)
  AUTHORS   Sands B, Burnaevskiy N, Yun SR, Crane MM, Kaeberlein M, Mendenhall A
  TITLE     A toolkit for DNA assembly, genome engineering and multicolor 
            imaging for C. elegans.
  JOURNAL   Transl Med Aging. 2018 Jan;2:1-10. doi: 10.1016/j.tma.2018.01.001. 
            Epub 2018 Apr 13.
  PUBMED    33693300
REFERENCE   2  (bases 1 to 8495)
  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: "Transl Med 
            Aging."; date: "2018-01"; volume: "2:1-10. doi"; pages: " 
            10.1016/j.tma.2018.01.001. Epub 2018 Apr 13"
COMMENT     Sequence Label: BSP179
FEATURES             Location/Qualifiers
     source          1..8495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    29..53
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(199..210)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2150..2167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2151..2167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2363..2379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2363..2379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(2386..2410)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3838..3858)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     terminator      complement(3886..4082)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(4088..4903)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4088..4903)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4811..4830
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(4913..4935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4924..4940
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4949..4954,4955..4970,4971..4976))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     rep_origin      5331..5919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5820..5839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      5982..7145
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    7168..7188
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 attccatgat ggtagcaaac tcacttcgtc aagtttgtac aaaaaagcag gcttgagcca
       61 atttatccaa gtccttgtaa aaagtattcg aaaattgtta acggataaaa tgtttattat
      121 aatatcaaaa acaatttgtc agttgaccac tttttgatat atttgacaga aaccgggatg
      181 aattggctca aagttagggc gcccttctat tacagggttt ctgataacaa accggttatt
      241 aactcccaac aagggatgat ttcaattcat catgctcaaa ttgacccaaa ttaagttaca
      301 tgacaaattc atcgcccttt tcaactcttc ttggctcatc atctgttatt ctgttcctat
      361 tctgtctgca ccctataccc ctttgcatac tctctcgtca ttcctttctt ggatagtgtc
      421 ttcttctccc cagctctgct acttctatga cttgcgcccg ctgcttttcc gctcgttcct
      481 ctcctctctg acgtcttctt ctgcttcttt tctccttcct ttttcccttc tcaatcttcc
      541 tctttttccc atttctgtca atcattcgaa gaagaagaag aagaagaccc tcattcattt
      601 catttttttt tctgtcggtg tgtgctgctc ggttaagagt gagctctcta ttccacgtct
      661 tcttcttttt cttcttcgat tcgaatcaat cactccacca aaacgcattc gtttttggga
      721 ttcaccccgc ggttcgcaat aggtttcttt ttcaaattat tagcgttata aatagaaaaa
      781 tggtggagtt tcaataaaaa tgataattta caaagtgtat tttgattaca tgtactcaaa
      841 aaggttgaaa tttttcatac cagttttccg gaaatccatc tgatatcatt atcgtatttt
      901 tctttttaaa aatgttttca aaaaaaaaca aaatatagct ggttatttgg caccctctaa
      961 ttaccatttt cctgtcacac cacactcttt tccttcttcc acttctttcc gttttcagcc
     1021 gcttccaacc aaaccgatat gaaagccgag caacaacgat cgattccacc cggctcggcg
     1081 accttcccgt cgcaggtgag actcagaaaa ctagagaaac cgctcaacta actcttgata
     1141 tccgatttca ttcttttctc ttttcttttt gttgaacttt tccatatttc cagatgccac
     1201 ggccaccacc aagcaccgaa caaggaatca caacggaatc ggagcttgcg aagaaagctc
     1261 aaatcactcc gaacgacgtt ttagcacttc cgggaatcac tcaaggtatc ttgctccctt
     1321 tggatttcag tttaaaacat aaatttagga ttcttatgct ccccatctgc gaacatctat
     1381 aacatcgagt tcaccaagtt ccaaatccgt gatctggaca ctgagcaagt gcttttcgag
     1441 atcgccaaac cggagaacga tcaggagaat gatgagtcgc cacaggagtc ggcaagatac
     1501 gtgcgttata gatttgctcc aaactttttg aaactcaaaa cggtcggagc aactgtggag
     1561 ttcaaagtag gagacatccc aatccatcat ttccgaatga tcgaacgtca cttcttcaaa
     1621 gatcgccttc tgaagtgttt tgactttgaa tttggattct gtattccgaa ctcacgaaac
     1681 aactgtgaac atatctatga gttccctcaa ctctctcaac aactcagtga gtcattattc
     1741 taaaaagtac aattcaaaag actaatctct ttttcagtgg acgacatgat caacaatcca
     1801 aacgagactc gttctgacag cttctatttc gtcgataaca aactcgtcat gcacaacaaa
     1861 gccgactact catatgatgc ataaatattt aatacaaaaa tgttctggat aattattctg
     1921 tcgaatagaa aaaaaactcc aaatgtgatt aaattccaat aattcctgtc tagtttgttc
     1981 cttccccttc cccttctcat gttcaatgca ttcctaagct tttcagttcc ccccttgttt
     2041 ctatattttt tcgctgtcct gtcacactcg ctaaaaacac taatcacacg gaaatctgtt
     2101 ttcaataaaa actccaactt taactcattt tcaatttcaa ctgcacgggt gtaaaacgac
     2161 ggccagtccg taatacgact cacttaaggc cttgactaga gggtaccaga gctcacctag
     2221 gtatctagaa ccggtgacgt caccatggga agcttcgtgg atccagatat cctgcaggaa
     2281 ttcctcgaga cgtacggtgc gcgcgatgca ttcgaagatc tgcccactag tgagtcgtat
     2341 tacgtagctt ggcgtaatca tggtcatagc tgtttcctgc ccgtgaccca gctttcttgt
     2401 acaaagtggt aagtgcaagt aagatcagtg tttgtttcga tatcagtctg tttcgtaacg
     2461 gtcttctgta taactacaaa aaagaataaa aaaccgtatc acaactgaga aaaaagacaa
     2521 aacttattga acagtgttcc tttcaaaaaa ttattttgta aaaaatcgag tttttctcca
     2581 aaataacgca cgcaatcaaa aacaagaaaa ctaacgtgtt ctcctgggcg tggaactcct
     2641 tatcaagatt accattattg tctgtcagaa ttgatatttc aattatttca tatctagaca
     2701 aacccgggtg tcttgcattt attagtactg tgagtttaca tacaaacgag gcttactcag
     2761 ataacataga gtcacctcta attagcaaaa acccatttct aattagctat ttcgaccagt
     2821 tctaaagtca ccgtaattag aaatggtttt tgtagctttt ctaattagaa atgaccccga
     2881 gataagtgaa atatggcatt ttctgggttt tctagtaaga aatgatcccg cacgtagtga
     2941 ttccgcgtta caagccaaat tcaaattaaa aaagtatatt acacaattca agttacatta
     3001 ttttcaggaa acaatggcga agagcaaagc agagattcaa aaggaggaaa gagacaacag
     3061 atatgaacga agacttcttg aaaagagaaa gagacatgag gtgagtaacc atgataataa
     3121 ataatcaagt aagggctata gatttataaa ttctggagtt aacagtttcg aaaaaaagta
     3181 gttttcatat gtgcgaagga atatcaaaaa taggggaata gcctaaacga gtgagaaaat
     3241 taaatgtttg ttattttttc aaaaatgtct gcagggccgt tatcctttca aatgcactct
     3301 atcaaacata gatcatatcc atcatatctt atccagaatt tatatcatat ccaagattga
     3361 gagcgtttca aggaaaatct caaacaacca tacaattttg tagatcgaca acaaggcctc
     3421 cacttcttca tcgatggcct ctgcagcacc tgcaccacca agtttgtatt tttcatatta
     3481 atataaagtg agccacgcat agtcgttttt aggatttttg ccagatcgat gtcgcgcaaa
     3541 cagactgttt ttttttcttt gaaagccgtg cgtgacatgg gccacggatt gtaacgccgc
     3601 gctggatgga taaaaattta ttcagttttt tagcctttac aaaatatggt tcgactactt
     3661 tctttatttt tacattttta taggtgctca ttgagctgag atattttggg tataatcgga
     3721 aaatcgctga ttttggtcat ttttctgcac taaagattac acaaaaagtt cagtatttag
     3781 attcattgtt gaaaattggg gatgatctac agatacatat cgccgtgttc aaaaacacaa
     3841 ctttattata catagttgat aattcgatgg tatcagctca ctcaaagtat agcgaccagc
     3901 attcacatac gattgacgca tgatattact ttctgcgcac ttaacttcgc atctgggcag
     3961 atgatgtcga ggcgaaaaaa aatataaatc acgctaacat ttgattaaaa tagaacaact
     4021 acaatataaa aaaactatac aaatgacaag ttcttgaaaa caagaatctt tttattgtca
     4081 gtactgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     4141 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     4201 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     4261 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     4321 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     4381 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     4441 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     4501 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     4561 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     4621 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     4681 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     4741 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     4801 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     4861 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cattttatgt ttcctgtgtg
     4921 aaattgttat ccgctcacaa ttccacagat tatacgagcc gatgattaat tgtcaacagc
     4981 tctttatgtt cggatgtgat gtgagaactg tatcctagca agattttaaa aggaagtata
     5041 tgaaagaaga acctcagtgg caaatcctaa ccttttatat ttctctacag gggcgcggcg
     5101 tggggacaat tcaacgcgtc tgtgagggga gcgtttccct gctcgcaggt ctgcagcgag
     5161 gagccgtaat ttttgcttcg cgccgtgcgg ccatcaaaat gtatggatgc aaatgattat
     5221 acatggggat gtatgggcta aatgtacggg cgacagtcac atcatgcccc tgagctgcgc
     5281 acgtcaagac tgtcaaggag ggtattctgg gcctccatgt cgggatcttc ttgagatcct
     5341 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5401 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5461 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5521 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5581 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5641 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5701 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5761 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5821 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5881 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5941 ttacggttcc tggccttttg ctggccgaaa agtgccacct gaacgaagca tctgtgcttc
     6001 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     6061 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     6121 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     6181 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     6241 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     6301 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     6361 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     6421 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     6481 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     6541 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     6601 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     6661 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     6721 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     6781 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     6841 tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt
     6901 tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt
     6961 ctagctagag aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct
     7021 tccgaaaatg caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct
     7081 gcgtgttgcc tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa
     7141 atgcgcgcca gggatccgcc aagctatcaa ctttgtatag aaaagttgtg tcggattatg
     7201 ggctcttctt ttatcctcta ctttctttct agaaaatgca aatgttgagt cagaaattgc
     7261 tgaatggagt acaactgcaa tgggtttagc ttatgtttct ggaacaatta ctggtggaat
     7321 tgtgattgaa aagtaaggtt atatttcatt tcaataaatg ttaccacagt ataacaaaaa
     7381 caagttacat tgttcagagt atgtctgaat gtattattat tattgtattt gtgcatcagt
     7441 actgtagttg tactgaagaa gggtttaaaa taatctcaac gtttttaaga cgagttagtg
     7501 agaatacggt cgcagttagg atgtttggac atagttttat tagataatcg tgaactaact
     7561 cactataaaa acccgctaac tcattgaata agatagtttt aaacgttcaa aatttgaaga
     7621 caaaattgta tgaataagtt tacagagttg gtcgacgcaa gcttctcctt ctcttcacat
     7681 ttctcaataa tttagctcta attgcattcg tattcttcgc aaaaatacgt attctgattg
     7741 acccgatgaa atacggatgc ctagttgcat tgatcatcta cggttatact tatgggtgag
     7801 gtttttcaaa tatattttga tttaatcgaa taatgaaatt tcagaactgg agttggtcca
     7861 atttcgtggt ttatatcctc tgaactggta cctcaaaaac atcgtagtat tgctcaatct
     7921 gtggcatatt caatgaatac tgtaatggtg gttatcagta catttacagt attacctcta
     7981 tattccgtta ttggtagtta tgctttcctt attctatatt caattccttc atttataagt
     8041 atgcttatat tatttcgcta cttaccggaa accaaaggac gagagattca tgatattgtg
     8101 aatgaactta aaaaaaaatg aaaacctttg aaaacgttca tacggccgaa attttacttg
     8161 atcgagccaa ttgaactttt gttaagactt acagaaaagt gagagatttt gttacaaaaa
     8221 agtgagaaaa aagcgaaaat gaacaggaaa aagaagaaaa atacacccga agtatcatat
     8281 aaaaattaaa aagattgttt gacctggcgg aacttttttg caaaaactct gaatatccat
     8341 ggcacatgct tcgtgcaaaa caggatccaa atgaatatca acaagagatt cttgaagacg
     8401 acgagccact tgctcagagc attgcttatc tcgaatgaga cccttgttaa aatcattttt
     8461 caaacattat atgacattgt cgaaatgtcc tcctg
//
