LOCUS       sequence-242751-        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    3..27
                     /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             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        214..232
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    233..257
                     /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             2013..2057
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2095..2113)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2109..2156
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2386..3201)
                     /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(2386..3201)
                     /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     3109..3128
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3202..3293)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3309..4058)
                     /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(3319..3336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3559..3578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4254..5336)
                     /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     5298..5317
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5337..5414)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5460..5476,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
      241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
      301 tgttcgtgtc cgatttccgc aaagagttct acgaggtggt ccagagccag agggtccttc
      361 tcttcgtggc ctcggacgtg gatgctctgt gtgcgtgcaa gatccttcag gccttgttcc
      421 agtgtgacca cgtgcaatat acgctggttc cagtttctgg gtggcaagaa cttgaaactg
      481 catttcttga gcataaagaa cagtttcatt attttattct cataaactgt ggagctaatg
      541 tagacctatt ggatattctt caacctgatg aagacactat attctttgtg tgtgacaccc
      601 ataggccagt caatgtcgtc aatgtataca acgataccca gatcaaatta ctcattaaac
      661 aagatgatga ccttgaagtt cccgcctatg aagacatctt cagggatgaa gaggaggatg
      721 aagagcattc aggaaatgac agtgatgggt cagagccttc tgagaagcgc acacggttag
      781 aagaggagat agtggagcaa accatgcgga ggaggcagcg gcgagagtgg gaggcccgga
      841 gaagagacat cctctttgac tacgagcagt atgaatatca tgggacatcg tcagccatgg
      901 tgatgtttga gctggcttgg atgctgtcca aggacctgaa tgacatgctg tggtgggcca
      961 tcgttggact aacagaccag tgggtgcaag acaagatcac tcaaatgaaa tacgtgactg
     1021 atgttggtgt cctgcagcgc cacgtttccc gccacaacca ccggaacgag gatgaggaga
     1081 acacactctc cgtggactgc acacggatct cctttgagta tgacctccgc ctggtgctct
     1141 accagcactg gtccctccat gacagcctgt gcaacaccag ctataccgca gccaggttca
     1201 agctgtggtc tgtgcatgga cagaagcggc tccaggagtt ccttgcagac atgggtcttc
     1261 ccctgaagca ggtgaagcag aagttccagg ccatggacat ctccttgaag gagaatttgc
     1321 gggaaatgat tgaagagtct gcaaataaat ttgggatgaa ggacatgcgc gtgcagactt
     1381 tcagcattca ttttgggttc aagcacaagt ttctggccag cgacgtggtc tttgccacca
     1441 tgtctttgat ggagagcccc gagaaggatg gctcagggac agatcacttc atccaggctc
     1501 tggacagcct ctccaggagt aacctggaca agctgtacca tggcctggaa ctcgccaaga
     1561 agcagctgcg agccacccag cagaccattg ccagctgcct ttgcaccaac ctcgtcatct
     1621 cccaggggcc tttcctgtac tgctctctca tggagggcac tccagatgtc atgctgttct
     1681 ctaggccggc atccctaagc ctgctcagca aacacctgct caagtccttt gtgtgttcga
     1741 caaagaaccg gcgctgcaaa ctgctgcccc tggtgatggc tgcccccctg agcatggagc
     1801 atggcacagt gaccgtggtg ggcatccccc cagagaccga cagctcggac aggaagaact
     1861 tttttgggag ggcgtttgag aaggcagcgg aaagcaccag ctcccggatg ctgcacaacc
     1921 attttgacct ctcagtaatt gagctgaaag ctgaggatcg gagcaagttt ctggacgcac
     1981 ttatttccct cctgtcctaa ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac
     2041 gccagcacat ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg
     2101 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2161 aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
     2221 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg acaagctgac
     2281 gaccgggtct ccgcaagtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2341 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     2401 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     2461 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     2521 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2581 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2641 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2701 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2761 aaatacgcgg tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     2821 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     2881 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     2941 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     3001 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     3061 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     3121 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     3181 ttcccgttga atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggcat
     3301 gcagcgctct tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     3361 agcggtgtca gctcactcaa aagcggtaat acggttatcc acagaatcag gggataaagc
     3421 cggaaagaac atgtgagcaa aaagcaaagc accggaagaa gccaacgccg caggcgtttt
     3481 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagc cagaggtggc
     3541 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3601 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3661 tggcgctttc tcatagctca cgctgttggt atctcagttc ggtgtaggtc gttcgctcca
     3721 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3781 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccattggta
     3841 actgatttag aggactttgt cttgaagtta tgcacctgtt aaggctaaac tgaaagaaca
     3901 gattttggtg agtgcggtcc tccaacccac ttaccttggt tcaaagagtt ggtagctcag
     3961 cgaaccttga gaaaaccacc gttggtagcg gtggtttttc tttatttatg agatgatgaa
     4021 tcaatcggtc tatcaagtca acgaacagct attccgttac tctagatttc agtgcaattt
     4081 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt
     4141 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     4201 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     4261 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4321 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     4381 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     4441 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     4501 ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac
     4561 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     4621 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     4681 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     4741 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     4801 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     4861 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     4921 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     4981 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     5041 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     5101 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     5161 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     5221 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     5281 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     5341 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     5401 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaatt
     5461 aatacgactc actata
//
