LOCUS       sequence-213752-        9239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cloning by HR in yeast, Universal MosSCI compatible, has unc54 
            terminator cloned.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9239)
  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 9239)
  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: BSP188
FEATURES             Location/Qualifiers
     source          1..9239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(147..158)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2098..2115
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2099..2115
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3055..3071)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3055..3071)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(3078..3102)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4530..4550)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     terminator      complement(4578..4774)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(4780..5595)
                     /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(4780..5595)
                     /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     5503..5522
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(5605..5627)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5616..5632
                     /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(5641..5646,5647..5662,5663..5668))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     rep_origin      6023..6611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6512..6531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      6674..7837
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    7860..7880
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    join(9216..9239,1)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgagccaat ttatccaagt ccttgtaaaa agtattcgaa aattgttaac ggataaaatg
       61 tttattataa tatcaaaaac aatttgtcag ttgaccactt tttgatatat ttgacagaaa
      121 ccgggatgaa ttggctcaaa gttagggcgc ccttctatta cagggtttct gataacaaac
      181 cggttattaa ctcccaacaa gggatgattt caattcatca tgctcaaatt gacccaaatt
      241 aagttacatg acaaattcat cgcccttttc aactcttctt ggctcatcat ctgttattct
      301 gttcctattc tgtctgcacc ctatacccct ttgcatactc tctcgtcatt cctttcttgg
      361 atagtgtctt cttctcccca gctctgctac ttctatgact tgcgcccgct gcttttccgc
      421 tcgttcctct cctctctgac gtcttcttct gcttcttttc tccttccttt ttcccttctc
      481 aatcttcctc tttttcccat ttctgtcaat cattcgaaga agaagaagaa gaagaccctc
      541 attcatttca tttttttttc tgtcggtgtg tgctgctcgg ttaagagtga gctctctatt
      601 ccacgtcttc ttctttttct tcttcgattc gaatcaatca ctccaccaaa acgcattcgt
      661 ttttgggatt caccccgcgg ttcgcaatag gtttcttttt caaattatta gcgttataaa
      721 tagaaaaatg gtggagtttc aataaaaatg ataatttaca aagtgtattt tgattacatg
      781 tactcaaaaa ggttgaaatt tttcatacca gttttccgga aatccatctg atatcattat
      841 cgtatttttc tttttaaaaa tgttttcaaa aaaaaacaaa atatagctgg ttatttggca
      901 ccctctaatt accattttcc tgtcacacca cactcttttc cttcttccac ttctttccgt
      961 tttcagccgc ttccaaccaa accgatatga aagccgagca acaacgatcg attccacccg
     1021 gctcggcgac cttcccgtcg caggtgagac tcagaaaact agagaaaccg ctcaactaac
     1081 tcttgatatc cgatttcatt cttttctctt ttctttttgt tgaacttttc catatttcca
     1141 gatgccacgg ccaccaccaa gcaccgaaca aggaatcaca acggaatcgg agcttgcgaa
     1201 gaaagctcaa atcactccga acgacgtttt agcacttccg ggaatcactc aaggtatctt
     1261 gctccctttg gatttcagtt taaaacataa atttaggatt cttatgctcc ccatctgcga
     1321 acatctataa catcgagttc accaagttcc aaatccgtga tctggacact gagcaagtgc
     1381 ttttcgagat cgccaaaccg gagaacgatc aggagaatga tgagtcgcca caggagtcgg
     1441 caagatacgt gcgttataga tttgctccaa actttttgaa actcaaaacg gtcggagcaa
     1501 ctgtggagtt caaagtagga gacatcccaa tccatcattt ccgaatgatc gaacgtcact
     1561 tcttcaaaga tcgccttctg aagtgttttg actttgaatt tggattctgt attccgaact
     1621 cacgaaacaa ctgtgaacat atctatgagt tccctcaact ctctcaacaa ctcagtgagt
     1681 cattattcta aaaagtacaa ttcaaaagac taatctcttt ttcagtggac gacatgatca
     1741 acaatccaaa cgagactcgt tctgacagct tctatttcgt cgataacaaa ctcgtcatgc
     1801 acaacaaagc cgactactca tatgatgcat aaatatttaa tacaaaaatg ttctggataa
     1861 ttattctgtc gaatagaaaa aaaactccaa atgtgattaa attccaataa ttcctgtcta
     1921 gtttgttcct tccccttccc cttctcatgt tcaatgcatt cctaagcttt tcagttcccc
     1981 ccttgtttct atattttttc gctgtcctgt cacactcgct aaaaacacta atcacacgga
     2041 aatctgtttt caataaaaac tccaacttta actcattttc aatttcaact gcacgggtgt
     2101 aaaacgacgg ccagtccgta atacgactca cttaaggcct tgactagagg gtaccagagc
     2161 tcacctaggt atctagaacc ggtgacgtca ccatgggaag cttcgtggat ccagatatcc
     2221 tgcaggaatt cctcgagacg tacggtgcgc gcgatgcatt cgaagatctg gccgctgtca
     2281 tcagacatct cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac
     2341 atgctctttc tccctgtgct cccaccccct atttttgtta ttatcaaaaa acttctctta
     2401 atttctttgt tttttagctt cttttaagtc acctctaaca atgaaattgt gtagattcaa
     2461 aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt gctccctccc cccattaata
     2521 ataattctat cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt ttttacttct
     2581 gataaatttt tttgaaacat catagaaaaa accgcacaca aaatacctta tcatatgtta
     2641 cgtttcagtt tatgaccgca atttttattt cttcgcacgt ctgggcctct catgacgtca
     2701 aatcatgctc atcgtgaaaa agttttggag tatttttgga atttttcaat caagtgaaag
     2761 tttatgaaat taattttcct gcttttgctt tttggggttt cccctattgt ttgtcaagat
     2821 ttcgaggacg gcgtttttct tgctaaaatc acaagtattg atgagcacga tgcaagaaag
     2881 atcggaagaa ggtttgggtt tgaggctcag tggaaggtga gtagaagttg ataatttgaa
     2941 agtggagtag tgtctatggg gtttttgcct taaatgacag aatacattcc caatatacca
     3001 aacataactg tttgcccact agtgagtcgt attacgtagc ttggcgtaat catggtcata
     3061 gctgtttcct gcccgtgacc cagctttctt gtacaaagtg gtaagtgcaa gtaagatcag
     3121 tgtttgtttc gatatcagtc tgtttcgtaa cggtcttctg tataactaca aaaaagaata
     3181 aaaaaccgta tcacaactga gaaaaaagac aaaacttatt gaacagtgtt cctttcaaaa
     3241 aattattttg taaaaaatcg agtttttctc caaaataacg cacgcaatca aaaacaagaa
     3301 aactaacgtg ttctcctggg cgtggaactc cttatcaaga ttaccattat tgtctgtcag
     3361 aattgatatt tcaattattt catatctaga caaacccggg tgtcttgcat ttattagtac
     3421 tgtgagttta catacaaacg aggcttactc agataacata gagtcacctc taattagcaa
     3481 aaacccattt ctaattagct atttcgacca gttctaaagt caccgtaatt agaaatggtt
     3541 tttgtagctt ttctaattag aaatgacccc gagataagtg aaatatggca ttttctgggt
     3601 tttctagtaa gaaatgatcc cgcacgtagt gattccgcgt tacaagccaa attcaaatta
     3661 aaaaagtata ttacacaatt caagttacat tattttcagg aaacaatggc gaagagcaaa
     3721 gcagagattc aaaaggagga aagagacaac agatatgaac gaagacttct tgaaaagaga
     3781 aagagacatg aggtgagtaa ccatgataat aaataatcaa gtaagggcta tagatttata
     3841 aattctggag ttaacagttt cgaaaaaaag tagttttcat atgtgcgaag gaatatcaaa
     3901 aataggggaa tagcctaaac gagtgagaaa attaaatgtt tgttattttt tcaaaaatgt
     3961 ctgcagggcc gttatccttt caaatgcact ctatcaaaca tagatcatat ccatcatatc
     4021 ttatccagaa tttatatcat atccaagatt gagagcgttt caaggaaaat ctcaaacaac
     4081 catacaattt tgtagatcga caacaaggcc tccacttctt catcgatggc ctctgcagca
     4141 cctgcaccac caagtttgta tttttcatat taatataaag tgagccacgc atagtcgttt
     4201 ttaggatttt tgccagatcg atgtcgcgca aacagactgt ttttttttct ttgaaagccg
     4261 tgcgtgacat gggccacgga ttgtaacgcc gcgctggatg gataaaaatt tattcagttt
     4321 tttagccttt acaaaatatg gttcgactac tttctttatt tttacatttt tataggtgct
     4381 cattgagctg agatattttg ggtataatcg gaaaatcgct gattttggtc atttttctgc
     4441 actaaagatt acacaaaaag ttcagtattt agattcattg ttgaaaattg gggatgatct
     4501 acagatacat atcgccgtgt tcaaaaacac aactttatta tacatagttg ataattcgat
     4561 ggtatcagct cactcaaagt atagcgacca gcattcacat acgattgacg catgatatta
     4621 ctttctgcgc acttaacttc gcatctgggc agatgatgtc gaggcgaaaa aaaatataaa
     4681 tcacgctaac atttgattaa aatagaacaa ctacaatata aaaaaactat acaaatgaca
     4741 agttcttgaa aacaagaatc tttttattgt cagtactgat tagaaaaact catcgagcat
     4801 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     4861 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     4921 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     4981 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     5041 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     5101 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     5161 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     5221 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     5281 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     5341 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     5401 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     5461 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     5521 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg
     5581 ttgaatatgg ctcattttat gtttcctgtg tgaaattgtt atccgctcac aattccacag
     5641 attatacgag ccgatgatta attgtcaaca gctctttatg ttcggatgtg atgtgagaac
     5701 tgtatcctag caagatttta aaaggaagta tatgaaagaa gaacctcagt ggcaaatcct
     5761 aaccttttat atttctctac aggggcgcgg cgtggggaca attcaacgcg tctgtgaggg
     5821 gagcgtttcc ctgctcgcag gtctgcagcg aggagccgta atttttgctt cgcgccgtgc
     5881 ggccatcaaa atgtatggat gcaaatgatt atacatgggg atgtatgggc taaatgtacg
     5941 ggcgacagtc acatcatgcc cctgagctgc gcacgtcaag actgtcaagg agggtattct
     6001 gggcctccat gtcgggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6061 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     6181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggccga
     6661 aaagtgccac ctgaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     6721 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     6781 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     6841 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     6901 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     6961 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     7021 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     7081 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     7141 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     7201 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     7261 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     7321 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     7381 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     7441 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     7501 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     7561 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg
     7621 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcggaata
     7681 ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca
     7741 tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg
     7801 agaagaacgg catagtgcgt gtttatgctt aaatgcgcgc cagggatccg ccaagctatc
     7861 aactttgtat agaaaagttg tgtcggatta tgggctcttc ttttatcctc tactttcttt
     7921 ctagaaaatg caaatgttga gtcagaaatt gctgaatgga gtacaactgc aatgggttta
     7981 gcttatgttt ctggaacaat tactggtgga attgtgattg aaaagtaagg ttatatttca
     8041 tttcaataaa tgttaccaca gtataacaaa aacaagttac attgttcaga gtatgtctga
     8101 atgtattatt attattgtat ttgtgcatca gtactgtagt tgtactgaag aagggtttaa
     8161 aataatctca acgtttttaa gacgagttag tgagaatacg gtcgcagtta ggatgtttgg
     8221 acatagtttt attagataat cgtgaactaa ctcactataa aaacccgcta actcattgaa
     8281 taagatagtt ttaaacgttc aaaatttgaa gacaaaattg tatgaataag tttacagagt
     8341 tggtcgacgc aagcttctcc ttctcttcac atttctcaat aatttagctc taattgcatt
     8401 cgtattcttc gcaaaaatac gtattctgat tgacccgatg aaatacggat gcctagttgc
     8461 attgatcatc tacggttata cttatgggtg aggtttttca aatatatttt gatttaatcg
     8521 aataatgaaa tttcagaact ggagttggtc caatttcgtg gtttatatcc tctgaactgg
     8581 tacctcaaaa acatcgtagt attgctcaat ctgtggcata ttcaatgaat actgtaatgg
     8641 tggttatcag tacatttaca gtattacctc tatattccgt tattggtagt tatgctttcc
     8701 ttattctata ttcaattcct tcatttataa gtatgcttat attatttcgc tacttaccgg
     8761 aaaccaaagg acgagagatt catgatattg tgaatgaact taaaaaaaaa tgaaaacctt
     8821 tgaaaacgtt catacggccg aaattttact tgatcgagcc aattgaactt ttgttaagac
     8881 ttacagaaaa gtgagagatt ttgttacaaa aaagtgagaa aaaagcgaaa atgaacagga
     8941 aaaagaagaa aaatacaccc gaagtatcat ataaaaatta aaaagattgt ttgacctggc
     9001 ggaacttttt tgcaaaaact ctgaatatcc atggcacatg cttcgtgcaa aacaggatcc
     9061 aaatgaatat caacaagaga ttcttgaaga cgacgagcca cttgctcaga gcattgctta
     9121 tctcgaatga gacccttgtt aaaatcattt ttcaaacatt atatgacatt gtcgaaatgt
     9181 cctcctgatt ccatgatggt agcaaactca cttcgtcaag tttgtacaaa aaagcaggc
//
