LOCUS       sequence-208482-        8494 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 8494)
  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 8494)
  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..8494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(146..157)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2097..2114
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2098..2114
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2310..2326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2310..2326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(2333..2357)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3785..3805)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     terminator      complement(3833..4029)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(4035..4850)
                     /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(4035..4850)
                     /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     4758..4777
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(4860..4882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4871..4887
                     /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(4896..4901,4902..4917,4918..4923))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     rep_origin      5278..5866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5767..5786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      5929..7091
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    7114..7134
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    8470..8494
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 tgagccaatt tatccaagtc cttgtaaaaa gtattcgaaa attgttaacg gataaaatgt
       61 ttattataat atcaaaaaca atttgtcagt tgaccacttt ttgatatatt tgacagaaac
      121 cgggatgaat tggctcaaag ttagggcgcc cttctattac agggtttctg ataacaaacc
      181 ggttattaac tcccaacaag ggatgatttc aattcatcat gctcaaattg acccaaatta
      241 agttacatga caaattcatc gcccttttca actcttcttg gctcatcatc tgttattctg
      301 ttcctattct gtctgcaccc tatacccctt tgcatactct ctcgtcattc ctttcttgga
      361 tagtgtcttc ttctccccag ctctgctact tctatgactt gcgcccgctg cttttccgct
      421 cgttcctctc ctctctgacg tcttcttctg cttcttttct ccttcctttt tcccttctca
      481 atcttcctct ttttcccatt tctgtcaatc attcgaagaa gaagaagaag aagaccctca
      541 ttcatttcat ttttttttct gtcggtgtgt gctgctcggt taagagtgag ctctctattc
      601 cacgtcttct tctttttctt cttcgattcg aatcaatcac tccaccaaaa cgcattcgtt
      661 tttgggattc accccgcggt tcgcaatagg tttctttttc aaattattag cgttataaat
      721 agaaaaatgg tggagtttca ataaaaatga taatttacaa agtgtatttt gattacatgt
      781 actcaaaaag gttgaaattt ttcataccag ttttccggaa atccatctga tatcattatc
      841 gtatttttct ttttaaaaat gttttcaaaa aaaaacaaaa tatagctggt tatttggcac
      901 cctctaatta ccattttcct gtcacaccac actcttttcc ttcttccact tctttccgtt
      961 ttcagccgct tccaaccaaa ccgatatgaa agccgagcaa caacgatcga ttccacccgg
     1021 ctcggcgacc ttcccgtcgc aggtgagact cagaaaacta gagaaaccgc tcaactaact
     1081 cttgatatcc gatttcattc ttttctcttt tctttttgtt gaacttttcc atatttccag
     1141 atgccacggc caccaccaag caccgaacaa ggaatcacaa cggaatcgga gcttgcgaag
     1201 aaagctcaaa tcactccgaa cgacgtttta gcacttccgg gaatcactca aggtatcttg
     1261 ctccctttgg atttcagttt aaaacataaa tttaggattc ttatgctccc catctgcgaa
     1321 catctataac atcgagttca ccaagttcca aatccgtgat ctggacactg agcaagtgct
     1381 tttcgagatc gccaaaccgg agaacgatca ggagaatgat gagtcgccac aggagtcggc
     1441 aagatacgtg cgttatagat ttgctccaaa ctttttgaaa ctcaaaacgg tcggagcaac
     1501 tgtggagttc aaagtaggag acatcccaat ccatcatttc cgaatgatcg aacgtcactt
     1561 cttcaaagat cgccttctga agtgttttga ctttgaattt ggattctgta ttccgaactc
     1621 acgaaacaac tgtgaacata tctatgagtt ccctcaactc tctcaacaac tcagtgagtc
     1681 attattctaa aaagtacaat tcaaaagact aatctctttt tcagtggacg acatgatcaa
     1741 caatccaaac gagactcgtt ctgacagctt ctatttcgtc gataacaaac tcgtcatgca
     1801 caacaaagcc gactactcat atgatgcata aatatttaat acaaaaatgt tctggataat
     1861 tattctgtcg aatagaaaaa aaactccaaa tgtgattaaa ttccaataat tcctgtctag
     1921 tttgttcctt ccccttcccc ttctcatgtt caatgcattc ctaagctttt cagttccccc
     1981 cttgtttcta tattttttcg ctgtcctgtc acactcgcta aaaacactaa tcacacggaa
     2041 atctgttttc aataaaaact ccaactttaa ctcattttca atttcaactg cacgggtgta
     2101 aaacgacggc cagtccgtaa tacgactcac ttaaggcctt gactagaggg taccagagct
     2161 cacctaggta tctagaaccg gtgacgtcac catgggaagc ttcgtggatc cagatatcct
     2221 gcaggaattc ctcgagacgt acggtgcgcg cgatgcattc gaagatctgc ccactagtga
     2281 gtcgtattac gtagcttggc gtaatcatgg tcatagctgt ttcctgcccg tgacccagct
     2341 ttcttgtaca aagtggtaag tgcaagtaag atcagtgttt gtttcgatat cagtctgttt
     2401 cgtaacggtc ttctgtataa ctacaaaaaa gaataaaaaa ccgtatcaca actgagaaaa
     2461 aagacaaaac ttattgaaca gtgttccttt caaaaaatta ttttgtaaaa aatcgagttt
     2521 ttctccaaaa taacgcacgc aatcaaaaac aagaaaacta acgtgttctc ctgggcgtgg
     2581 aactccttat caagattacc attattgtct gtcagaattg atatttcaat tatttcatat
     2641 ctagacaaac ccgggtgtct tgcatttatt agtactgtga gtttacatac aaacgaggct
     2701 tactcagata acatagagtc acctctaatt agcaaaaacc catttctaat tagctatttc
     2761 gaccagttct aaagtcaccg taattagaaa tggtttttgt agcttttcta attagaaatg
     2821 accccgagat aagtgaaata tggcattttc tgggttttct agtaagaaat gatcccgcac
     2881 gtagtgattc cgcgttacaa gccaaattca aattaaaaaa gtatattaca caattcaagt
     2941 tacattattt tcaggaaaca atggcgaaga gcaaagcaga gattcaaaag gaggaaagag
     3001 acaacagata tgaacgaaga cttcttgaaa agagaaagag acatgaggtg agtaaccatg
     3061 ataataaata atcaagtaag ggctatagat ttataaattc tggagttaac agtttcgaaa
     3121 aaaagtagtt ttcatatgtg cgaaggaata tcaaaaatag gggaatagcc taaacgagtg
     3181 agaaaattaa atgtttgtta ttttttcaaa aatgtctgca gggccgttat cctttcaaat
     3241 gcactctatc aaacatagat catatccatc atatcttatc cagaatttat atcatatcca
     3301 agattgagag cgtttcaagg aaaatctcaa acaaccatac aattttgtag atcgacaaca
     3361 aggcctccac ttcttcatcg atggcctctg cagcacctgc accaccaagt ttgtattttt
     3421 catattaata taaagtgagc cacgcatagt cgtttttagg atttttgcca gatcgatgtc
     3481 gcgcaaacag actgtttttt tttctttgaa agccgtgcgt gacatgggcc acggattgta
     3541 acgccgcgct ggatggataa aaatttattc agttttttag cctttacaaa atatggttcg
     3601 actactttct ttatttttac atttttatag gtgctcattg agctgagata ttttgggtat
     3661 aatcggaaaa tcgctgattt tggtcatttt tctgcactaa agattacaca aaaagttcag
     3721 tatttagatt cattgttgaa aattggggat gatctacaga tacatatcgc cgtgttcaaa
     3781 aacacaactt tattatacat agttgataat tcgatggtat cagctcactc aaagtatagc
     3841 gaccagcatt cacatacgat tgacgcatga tattactttc tgcgcactta acttcgcatc
     3901 tgggcagatg atgtcgaggc gaaaaaaaat ataaatcacg ctaacatttg attaaaatag
     3961 aacaactaca atataaaaaa actatacaaa tgacaagttc ttgaaaacaa gaatcttttt
     4021 attgtcagta ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     4081 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     4141 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     4201 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     4261 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     4321 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     4381 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     4441 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     4501 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     4561 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     4621 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     4681 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     4741 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     4801 gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat tttatgtttc
     4861 ctgtgtgaaa ttgttatccg ctcacaattc cacagattat acgagccgat gattaattgt
     4921 caacagctct ttatgttcgg atgtgatgtg agaactgtat cctagcaaga ttttaaaagg
     4981 aagtatatga aagaagaacc tcagtggcaa atcctaacct tttatatttc tctacagggg
     5041 cgcggcgtgg ggacaattca acgcgtctgt gaggggagcg tttccctgct cgcaggtctg
     5101 cagcgaggag ccgtaatttt tgcttcgcgc cgtgcggcca tcaaaatgta tggatgcaaa
     5161 tgattataca tggggatgta tgggctaaat gtacgggcga cagtcacatc atgcccctga
     5221 gctgcgcacg tcaagactgt caaggagggt attctgggcc tccatgtcgg gatcttcttg
     5281 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5341 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5401 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5461 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5521 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5581 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5641 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5701 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5761 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5821 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5881 ggccttttta cggttcctgg ccttttgctg gccgaaaagt gccacctgac gaagcatctg
     5941 tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     6001 tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa
     6061 tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     6121 gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca
     6181 aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa
     6241 caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata
     6301 actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc
     6361 ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg
     6421 tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat
     6481 actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg
     6541 gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt
     6601 ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag taatactaga
     6661 gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga
     6721 tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa
     6781 tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg
     6841 gttttttgaa agtgcgtctt cagagcgctt ttggttttca aaagcgctct gaagttccta
     6901 tactttctag ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg
     6961 agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca cgtcgcacct
     7021 atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat
     7081 gcttaaatgc gcgccaggga tccgccaagc tatcaacttt gtatagaaaa gttgtgtcgg
     7141 attatgggct cttcttttat cctctacttt ctttctagaa aatgcaaatg ttgagtcaga
     7201 aattgctgaa tggagtacaa ctgcaatggg tttagcttat gtttctggaa caattactgg
     7261 tggaattgtg attgaaaagt aaggttatat ttcatttcaa taaatgttac cacagtataa
     7321 caaaaacaag ttacattgtt cagagtatgt ctgaatgtat tattattatt gtatttgtgc
     7381 atcagtactg tagttgtact gaagaagggt ttaaaataat ctcaacgttt ttaagacgag
     7441 ttagtgagaa tacggtcgca gttaggatgt ttggacatag ttttattaga taatcgtgaa
     7501 ctaactcact ataaaaaccc gctaactcat tgaataagat agttttaaac gttcaaaatt
     7561 tgaagacaaa attgtatgaa taagtttaca gagttggtcg acgcaagctt ctccttctct
     7621 tcacatttct caataattta gctctaattg cattcgtatt cttcgcaaaa atacgtattc
     7681 tgattgaccc gatgaaatac ggatgcctag ttgcattgat catctacggt tatacttatg
     7741 ggtgaggttt ttcaaatata ttttgattta atcgaataat gaaatttcag aactggagtt
     7801 ggtccaattt cgtggtttat atcctctgaa ctggtacctc aaaaacatcg tagtattgct
     7861 caatctgtgg catattcaat gaatactgta atggtggtta tcagtacatt tacagtatta
     7921 cctctatatt ccgttattgg tagttatgct ttccttattc tatattcaat tccttcattt
     7981 ataagtatgc ttatattatt tcgctactta ccggaaacca aaggacgaga gattcatgat
     8041 attgtgaatg aacttaaaaa aaaatgaaaa cctttgaaaa cgttcatacg gccgaaattt
     8101 tacttgatcg agccaattga acttttgtta agacttacag aaaagtgaga gattttgtta
     8161 caaaaaagtg agaaaaaagc gaaaatgaac aggaaaaaga agaaaaatac acccgaagta
     8221 tcatataaaa attaaaaaga ttgtttgacc tggcggaact tttttgcaaa aactctgaat
     8281 atccatggca catgcttcgt gcaaaacagg atccaaatga atatcaacaa gagattcttg
     8341 aagacgacga gccacttgct cagagcattg cttatctcga atgagaccct tgttaaaatc
     8401 atttttcaaa cattatatga cattgtcgaa atgtcctcct gattccatga tggtagcaaa
     8461 ctcacttcgt caagtttgta caaaaaagca ggct
//
