LOCUS       sequence-245619-        5476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Cdc45 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5476)
  AUTHORS   Simon AC, Sannino V, Costanzo V, Pellegrini L
  TITLE     Structure of human Cdc45 and implications for CMG helicase function.
  JOURNAL   Nat Commun. 2016 May 18;7:11638. doi: 10.1038/ncomms11638.
  PUBMED    27189187
REFERENCE   2  (bases 1 to 5476)
  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: "Nat 
            Commun."; date: "2016-05-18"; volume: "7:11638. doi"; pages: " 
            10.1038/ncomms11638"
COMMENT     Sequence Label: pRSFDuet1-Cdc45
FEATURES             Location/Qualifiers
     source          1..5476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             81..98
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        212..230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    231..255
                     /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             2011..2055
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2093..2111)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2107..2154
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2384..3199)
                     /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(2384..3199)
                     /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     3107..3126
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3200..3291)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3307..4056)
                     /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(3317..3334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3557..3576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4252..5334)
                     /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     5296..5315
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5335..5412)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        5458..5476
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctgt agaaataatt ttgtttaact ttaataagga
       61 gatataccat gggcagcagc catcaccatc atcaccacag ccaggatccg aattcgagct
      121 cggcgcgcct gcaggtcgac aagcttgcgg ccgcataatg cttaagtcga acagaaagta
      181 atcgtattgt acacggccgc ataatcgaaa ttaatacgac tcactatagg ggaattgtga
      241 gcggataaca attccccatc ttagtatatt agttaagtat aagaaggaga tatacatatg
      301 ttcgtgtccg atttccgcaa agagttctac gaggtggtcc agagccagag ggtccttctc
      361 ttcgtggcct cggacgtgga tgctctgtgt gcgtgcaaga tccttcaggc cttgttccag
      421 tgtgaccacg tgcaatatac gctggttcca gtttctgggt ggcaagaact tgaaactgca
      481 tttcttgagc ataaagaaca gtttcattat tttattctca taaactgtgg agctaatgta
      541 gacctattgg atattcttca acctgatgaa gacactatat tctttgtgtg tgacacccat
      601 aggccagtca atgtcgtcaa tgtatacaac gatacccaga tcaaattact cattaaacaa
      661 gatgatgacc ttgaagttcc cgcctatgaa gacatcttca gggatgaaga ggaggatgaa
      721 gagcattcag gaaatgacag tgatgggtca gagccttctg agaagcgcac acggttagaa
      781 gaggagatag tggagcaaac catgcggagg aggcagcggc gagagtggga ggcccggaga
      841 agagacatcc tctttgacta cgagcagtat gaatatcatg ggacatcgtc agccatggtg
      901 atgtttgagc tggcttggat gctgtccaag gacctgaatg acatgctgtg gtgggccatc
      961 gttggactaa cagaccagtg ggtgcaagac aagatcactc aaatgaaata cgtgactgat
     1021 gttggtgtcc tgcagcgcca cgtttcccgc cacaaccacc ggaacgagga tgaggagaac
     1081 acactctccg tggactgcac acggatctcc tttgagtatg acctccgcct ggtgctctac
     1141 cagcactggt ccctccatga cagcctgtgc aacaccagct ataccgcagc caggttcaag
     1201 ctgtggtctg tgcatggaca gaagcggctc caggagttcc ttgcagacat gggtcttccc
     1261 ctgaagcagg tgaagcagaa gttccaggcc atggacatct ccttgaagga gaatttgcgg
     1321 gaaatgattg aagagtctgc aaataaattt gggatgaagg acatgcgcgt gcagactttc
     1381 agcattcatt ttgggttcaa gcacaagttt ctggccagcg acgtggtctt tgccaccatg
     1441 tctttgatgg agagccccga gaaggatggc tcagggacag atcacttcat ccaggctctg
     1501 gacagcctct ccaggagtaa cctggacaag ctgtaccatg gcctggaact cgccaagaag
     1561 cagctgcgag ccacccagca gaccattgcc agctgccttt gcaccaacct cgtcatctcc
     1621 caggggcctt tcctgtactg ctctctcatg gagggcactc cagatgtcat gctgttctct
     1681 aggccggcat ccctaagcct gctcagcaaa cacctgctca agtcctttgt gtgttcgaca
     1741 aagaaccggc gctgcaaact gctgcccctg gtgatggctg cccccctgag catggagcat
     1801 ggcacagtga ccgtggtggg catcccccca gagaccgaca gctcggacag gaagaacttt
     1861 tttgggaggg cgtttgagaa ggcagcggaa agcaccagct cccggatgct gcacaaccat
     1921 tttgacctct cagtaattga gctgaaagct gaggatcgga gcaagtttct ggacgcactt
     1981 atttccctcc tgtcctaact cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc
     2041 cagcacatgg actcgtctac tagcgcagct taattaacct aggctgctgc caccgctgag
     2101 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     2161 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     2221 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga
     2281 ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2341 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     2401 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     2461 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     2521 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     2581 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     2641 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     2701 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     2761 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2821 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2881 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2941 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     3001 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     3061 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     3121 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     3181 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc
     3301 agcgctcttc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     3361 cggtgtcagc tcactcaaaa gcggtaatac ggttatccac agaatcaggg gataaagccg
     3421 gaaagaacat gtgagcaaaa agcaaagcac cggaagaagc caacgccgca ggcgtttttc
     3481 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagcca gaggtggcga
     3541 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3601 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3661 gcgctttctc atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3721 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3781 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cattggtaac
     3841 tgatttagag gactttgtct tgaagttatg cacctgttaa ggctaaactg aaagaacaga
     3901 ttttggtgag tgcggtcctc caacccactt accttggttc aaagagttgg tagctcagcg
     3961 aaccttgaga aaaccaccgt tggtagcggt ggtttttctt tatttatgag atgatgaatc
     4021 aatcggtcta tcaagtcaac gaacagctat tccgttactc tagatttcag tgcaatttat
     4081 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta
     4141 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     4201 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     4261 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4321 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     4381 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     4441 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     4501 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     4561 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     4621 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     4681 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     4741 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     4801 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     4861 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     4921 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     4981 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     5041 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     5101 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     5161 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     5221 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     5281 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     5341 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     5401 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaattaa
     5461 tacgactcac tatagg
//
