LOCUS       sequence-208485-        9238 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 9238)
  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 9238)
  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..9238
                     /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(3054..3070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3054..3070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(3077..3101)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4529..4549)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     terminator      complement(4577..4773)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(4779..5594)
                     /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(4779..5594)
                     /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     5502..5521
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(5604..5626)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5615..5631
                     /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(5640..5645,5646..5661,5662..5667))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     rep_origin      6022..6610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6511..6530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      6673..7835
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    7858..7878
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    9214..9238
                     /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 gaagatctgg 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 tttcccacta gtgagtcgta ttacgtagct tggcgtaatc atggtcatag
     3061 ctgtttcctg cccgtgaccc agctttcttg tacaaagtgg taagtgcaag taagatcagt
     3121 gtttgtttcg atatcagtct gtttcgtaac ggtcttctgt ataactacaa aaaagaataa
     3181 aaaaccgtat cacaactgag aaaaaagaca aaacttattg aacagtgttc ctttcaaaaa
     3241 attattttgt aaaaaatcga gtttttctcc aaaataacgc acgcaatcaa aaacaagaaa
     3301 actaacgtgt tctcctgggc gtggaactcc ttatcaagat taccattatt gtctgtcaga
     3361 attgatattt caattatttc atatctagac aaacccgggt gtcttgcatt tattagtact
     3421 gtgagtttac atacaaacga ggcttactca gataacatag agtcacctct aattagcaaa
     3481 aacccatttc taattagcta tttcgaccag ttctaaagtc accgtaatta gaaatggttt
     3541 ttgtagcttt tctaattaga aatgaccccg agataagtga aatatggcat tttctgggtt
     3601 ttctagtaag aaatgatccc gcacgtagtg attccgcgtt acaagccaaa ttcaaattaa
     3661 aaaagtatat tacacaattc aagttacatt attttcagga aacaatggcg aagagcaaag
     3721 cagagattca aaaggaggaa agagacaaca gatatgaacg aagacttctt gaaaagagaa
     3781 agagacatga ggtgagtaac catgataata aataatcaag taagggctat agatttataa
     3841 attctggagt taacagtttc gaaaaaaagt agttttcata tgtgcgaagg aatatcaaaa
     3901 ataggggaat agcctaaacg agtgagaaaa ttaaatgttt gttatttttt caaaaatgtc
     3961 tgcagggccg ttatcctttc aaatgcactc tatcaaacat agatcatatc catcatatct
     4021 tatccagaat ttatatcata tccaagattg agagcgtttc aaggaaaatc tcaaacaacc
     4081 atacaatttt gtagatcgac aacaaggcct ccacttcttc atcgatggcc tctgcagcac
     4141 ctgcaccacc aagtttgtat ttttcatatt aatataaagt gagccacgca tagtcgtttt
     4201 taggattttt gccagatcga tgtcgcgcaa acagactgtt tttttttctt tgaaagccgt
     4261 gcgtgacatg ggccacggat tgtaacgccg cgctggatgg ataaaaattt attcagtttt
     4321 ttagccttta caaaatatgg ttcgactact ttctttattt ttacattttt ataggtgctc
     4381 attgagctga gatattttgg gtataatcgg aaaatcgctg attttggtca tttttctgca
     4441 ctaaagatta cacaaaaagt tcagtattta gattcattgt tgaaaattgg ggatgatcta
     4501 cagatacata tcgccgtgtt caaaaacaca actttattat acatagttga taattcgatg
     4561 gtatcagctc actcaaagta tagcgaccag cattcacata cgattgacgc atgatattac
     4621 tttctgcgca cttaacttcg catctgggca gatgatgtcg aggcgaaaaa aaatataaat
     4681 cacgctaaca tttgattaaa atagaacaac tacaatataa aaaaactata caaatgacaa
     4741 gttcttgaaa acaagaatct ttttattgtc agtactgatt agaaaaactc atcgagcatc
     4801 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     4861 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     4921 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     4981 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     5041 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     5101 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     5161 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     5221 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     5281 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     5341 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     5401 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     5461 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     5521 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt
     5581 tgaatatggc tcattttatg tttcctgtgt gaaattgtta tccgctcaca attccacaga
     5641 ttatacgagc cgatgattaa ttgtcaacag ctctttatgt tcggatgtga tgtgagaact
     5701 gtatcctagc aagattttaa aaggaagtat atgaaagaag aacctcagtg gcaaatccta
     5761 accttttata tttctctaca ggggcgcggc gtggggacaa ttcaacgcgt ctgtgagggg
     5821 agcgtttccc tgctcgcagg tctgcagcga ggagccgtaa tttttgcttc gcgccgtgcg
     5881 gccatcaaaa tgtatggatg caaatgatta tacatgggga tgtatgggct aaatgtacgg
     5941 gcgacagtca catcatgccc ctgagctgcg cacgtcaaga ctgtcaagga gggtattctg
     6001 ggcctccatg tcgggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6061 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     6181 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccgaa
     6661 aagtgccacc tgacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     6721 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     6781 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     6841 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     6901 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     6961 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     7021 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     7081 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     7141 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     7201 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     7261 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     7321 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     7381 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     7441 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     7501 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
     7561 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt
     7621 ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact tcggaatagg
     7681 aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata
     7741 cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag
     7801 aagaacggca tagtgcgtgt ttatgcttaa atgcgcgcca gggatccgcc aagctatcaa
     7861 ctttgtatag aaaagttgtg tcggattatg ggctcttctt ttatcctcta ctttctttct
     7921 agaaaatgca aatgttgagt cagaaattgc tgaatggagt acaactgcaa tgggtttagc
     7981 ttatgtttct ggaacaatta ctggtggaat tgtgattgaa aagtaaggtt atatttcatt
     8041 tcaataaatg ttaccacagt ataacaaaaa caagttacat tgttcagagt atgtctgaat
     8101 gtattattat tattgtattt gtgcatcagt actgtagttg tactgaagaa gggtttaaaa
     8161 taatctcaac gtttttaaga cgagttagtg agaatacggt cgcagttagg atgtttggac
     8221 atagttttat tagataatcg tgaactaact cactataaaa acccgctaac tcattgaata
     8281 agatagtttt aaacgttcaa aatttgaaga caaaattgta tgaataagtt tacagagttg
     8341 gtcgacgcaa gcttctcctt ctcttcacat ttctcaataa tttagctcta attgcattcg
     8401 tattcttcgc aaaaatacgt attctgattg acccgatgaa atacggatgc ctagttgcat
     8461 tgatcatcta cggttatact tatgggtgag gtttttcaaa tatattttga tttaatcgaa
     8521 taatgaaatt tcagaactgg agttggtcca atttcgtggt ttatatcctc tgaactggta
     8581 cctcaaaaac atcgtagtat tgctcaatct gtggcatatt caatgaatac tgtaatggtg
     8641 gttatcagta catttacagt attacctcta tattccgtta ttggtagtta tgctttcctt
     8701 attctatatt caattccttc atttataagt atgcttatat tatttcgcta cttaccggaa
     8761 accaaaggac gagagattca tgatattgtg aatgaactta aaaaaaaatg aaaacctttg
     8821 aaaacgttca tacggccgaa attttacttg atcgagccaa ttgaactttt gttaagactt
     8881 acagaaaagt gagagatttt gttacaaaaa agtgagaaaa aagcgaaaat gaacaggaaa
     8941 aagaagaaaa atacacccga agtatcatat aaaaattaaa aagattgttt gacctggcgg
     9001 aacttttttg caaaaactct gaatatccat ggcacatgct tcgtgcaaaa caggatccaa
     9061 atgaatatca acaagagatt cttgaagacg acgagccact tgctcagagc attgcttatc
     9121 tcgaatgaga cccttgttaa aatcattttt caaacattat atgacattgt cgaaatgtcc
     9181 tcctgattcc atgatggtag caaactcact tcgtcaagtt tgtacaaaaa agcaggct
//
