LOCUS       sequence-420376-        5268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human Cdc45 C-terminally fused to 6xHis-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5268)
  AUTHORS   Szambowska A, Tessmer I, Kursula P, Usskilat C, Prus P, Pospiech H, 
            Grosse F
  TITLE     DNA binding properties of human Cdc45 suggest a function as 
            molecular wedge for DNA unwinding.
  JOURNAL   Nucleic Acids Res. 2014 Feb;42(4):2308-19. doi: 10.1093/nar/gkt1217.
            Epub 2013 Nov 28.
  PUBMED    24293646
REFERENCE   2  (bases 1 to 5268)
  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"; doi: 
            "10.1093/nar/gkt1217"; journalName: "Nucleic Acids Res"; date: 
            "2014-02"; volume: "42"; issue: "4"; pages: "2308-19"
COMMENT     Sequence Label: pRSFDuet-1-Cdc45-6His
FEATURES             Location/Qualifiers
     source          1..5268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    18..42
                     /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)."
     CDS             1787..1804
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1820..1864
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1902..1920)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1916..1963
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2193..3008)
                     /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(2193..3008)
                     /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     2916..2935
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3009..3100)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3116..3865)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3126..3143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3366..3385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4061..5143)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5105..5124
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5144..5221)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5267..5268,1..18)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5267..5268,1..17)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atacgactca ctatagggga attgtgagcg gataacaatt cccctgtaga aataattttg
       61 tttaacttta ataaggagat ataccatgtt cgtgtccgat ttccgcaaag agttctacga
      121 ggtggtccag agccagaggg tccttctctt cgtggcctcg gacgtggatg ctctgtgtgc
      181 gtgcaagatc cttcaggcct tgttccagtg tgaccacgtg caatatacgc tggttccagt
      241 ttctgggtgg caagaacttg aaactgcatt tcttgagcat aaagaacagt ttcattattt
      301 tattctcata aactgtggag ctaatgtaga cctattggat attcttcaac ctgatgaaga
      361 cactatattc tttgtgtgtg acacccatag gccagtcaat gtcgtcaatg tatacaacga
      421 tacccagatc aaattactca ttaaacaaga tgatgacctt gaagttcccg cctatgaaga
      481 catcttcagg gatgaagagg aggatgaaga gcattcagga aatgacagtg atgggtcaga
      541 gccttctgag aagcgcacac ggttagaaga ggagatagtg gagcaaacca tgcggaggag
      601 gcagcggcga gagtgggagg cccggagaag agacatcctc tttgactacg agcagtatga
      661 atatcatggg acatcgtcag ccatggtgat gtttgagctg gcttggatgc tgtccaagga
      721 cctgaatgac atgctgtggt gggccatcgt tggactaaca gaccagtggg tgcaagacaa
      781 gatcactcaa atgaaatacg tgactgatgt tggtgtcctg cagcgccacg tttcccgcca
      841 caaccaccgg aacgaggatg aggagaacac actctccgtg gactgcacac ggatctcctt
      901 tgagtatgac ctccgcctgg tgctctacca gcactggtcc ctccatgaca gcctgtgcaa
      961 caccagctat accgcagcca ggttcaagct gtggtctgtg catggacaga agcggctcca
     1021 ggagttcctt gcagacatgg gtcttcccct gaagcaggtg aagcagaagt tccaggccat
     1081 ggacatctcc ttgaaggaga atttgcggga aatgattgaa gagtctgcaa ataaatttgg
     1141 gatgaaggac atgcgcgtgc agactttcag cattcatttt gggttcaagc acaagtttct
     1201 ggccagcgac gtggtctttg ccaccatgtc tttgatggag agccccgaga aggatggctc
     1261 agggacagat cacttcatcc aggctctgga cagcctctcc aggagtaacc tggacaagct
     1321 gtaccatggc ctggaactcg ccaagaagca gctgcgagcc acccagcaga ccattgccag
     1381 ctgcctttgc accaacctcg tcatctccca ggggcctttc ctgtactgct ctctcatgga
     1441 gggcactcca gatgtcatgc tgttctctag gccggcatcc ctaagcctgc tcagcaaaca
     1501 cctgctcaag tcctttgtgt gttcgacaaa gaaccggcgc tgcaaactgc tgcccctggt
     1561 gatggctgcc cccctgagca tggagcatgg cacagtgacc gtggtgggca tccccccaga
     1621 gaccgacagc tcggacagga agaacttttt tgggagggcg tttgagaagg cagcggaaag
     1681 caccagctcc cggatgctgc acaaccattt tgacctctca gtaattgagc tgaaagctga
     1741 ggatcggagc aagtttctgg acgcacttat ttccctcctg tccggtcatc accatcacca
     1801 tcactaactc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga
     1861 ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc aataactagc
     1921 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt
     1981 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac
     2041 ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac cgggtctccg
     2101 caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2161 attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga
     2221 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     2281 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     2341 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     2401 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     2461 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     2521 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcggtcg
     2581 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     2641 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     2701 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     2761 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     2821 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     2881 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     2941 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     3001 tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca gcgctcttcc
     3121 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggtgtcagct
     3181 cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg aaagaacatg
     3241 tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc ataggctccg
     3301 cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa acccgacagg
     3361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3481 tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3601 caacccggta agacacgact tatcgccact ggcagcagcc attggtaact gatttagagg
     3661 actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat tttggtgagt
     3721 gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga accttgagaa
     3781 aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca atcggtctat
     3841 caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc tcttcaaatg
     3901 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg
     3961 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     4021 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4081 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4141 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     4201 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     4261 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     4321 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
     4381 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     4441 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     4501 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4561 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4621 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4681 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4741 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     4801 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     4861 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     4921 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     4981 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     5041 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     5101 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     5161 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     5221 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaatta
//
