LOCUS       sequence-225637-        8379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human MCM3 and human MCM5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8379)
  AUTHORS   Hesketh EL, Parker-Manuel RP, Chaban Y, Satti R, Coverley D, Orlova 
            EV, Chong JP
  TITLE     DNA induces conformational changes in a recombinant human 
            minichromosome maintenance complex.
  JOURNAL   J Biol Chem. 2015 Mar 20;290(12):7973-9. doi: 
            10.1074/jbc.M114.622738. Epub 2015 Feb 3.
  PUBMED    25648893
REFERENCE   2  (bases 1 to 8379)
  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: 
            "10.1074/jbc.M114.622738"; journalName: "J Biol Chem"; date: 
            "2015-03-20- 20"; volume: "290"; issue: "12"; pages: "7973-9"
COMMENT     Sequence Label: pRSF-MCM3/5
FEATURES             Location/Qualifiers
     source          1..8379
                     /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             77..94
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             98..112
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     promoter        2351..2369
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2370..2394
                     /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)."
     RBS             2409..2431
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4916..4960
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4998..5016)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5012..5059
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5289..6104)
                     /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(5289..6104)
                     /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     6012..6031
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(6105..6196)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6212..6961)
                     /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(6222..6239)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6462..6481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7157..8239)
                     /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     8201..8220
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8240..8317)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(8363..8379,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcacatc accaccacca tcacgtggat gacgacgaca agatgtcggg
      121 attcgacgat cctggcattt tctacagcga cagcttcggg ggcgacgccc aggccgacga
      181 ggggcaggcc cgcaaatcgc agctgcagag gcgcttcaag gagttcctgc ggcagtaccg
      241 agtgggcacc gaccgcacgg gcttcacctt caaatacagg gatgaactca agcggcatta
      301 caacctgggg gagtactgga ttgaggtgga gatggaggat ctggccagct ttgatgagga
      361 cctggccgac tacttgtaca agcagccagc cgagcacctg cagctgctgg aggaagctgc
      421 caaggaggta gctgatgagg tgacccggcc ccggccttct ggggaggagg tgctccagga
      481 catccaggtc atgctcaagt cggacgccag cccttccagc attcgtagcc tgaagtcgga
      541 catgatgtca cacctggtga agatccctgg catcatcatc gcggcctctg cggtccgtgc
      601 caaggccacc cgcatctcta tccagtgccg cagctgccgc aacaccctca gcaacattgc
      661 catgcgccct ggcctcgagg gctatgccct gcccaggaag tgcaacacag atcaggctgg
      721 gcgccccaaa tgcccattgg acccgtactt catcatgccc gacaaatgca aatgcgtgga
      781 cttccagacc ctgaagctgc aggagctgcc tgatgcagtc ccccacgggg agatgcccag
      841 acacatgcag ctctactgcg acaggtacct gtgtgacaag gtcgtccctg ggaacagggt
      901 taccatcatg ggcatctact ccatcaagaa gtttggcctg actaccagca ggggccgtga
      961 cagggtgggc gtgggcatcc gaagctccta catccgtgtc ctgggcatcc aggtggacac
     1021 agatggctct ggccgcagct ttgctggggc cgtgagcccc caggaggagg aggagttccg
     1081 tcgcctggct gccctcccaa atgtctatga ggtcatctcc aagagcatcg ccccctccat
     1141 ctttgggggc acagacatga agaaggccat tgcctgcctg ctctttgggg gctcccgaaa
     1201 gaggctccct gatggactta ctcgccgagg agacatcaac ctgctgatgc taggggaccc
     1261 tgggacagcc aagtcccagc ttctgaagtt tgtggagaag tgttctccca ttggggtata
     1321 cacgtctggg aaaggcagca gcgcagctgg actgacagcc tcggtgatga gggacccttc
     1381 gtcccggaat ttcatcatgg agggcggagc catggtcctg gccgatggtg gggtcgtctg
     1441 tattgacgag tttgacaaga tgcgagaaga tgaccgtgtg gcaatccacg aagccatgga
     1501 gcagcagacc atctctatcg ccaaggctgg gatcaccacc accctgaact cccgctgctc
     1561 cgtcctggct gctgccaact cagtgttcgg ccgctgggat gagacgaagg gggaggacaa
     1621 cattgacttc atgcccacca tcttgtcgcg cttcgacatg atcttcatcg tcaaggatga
     1681 gcacaatgag gagagggatg tgatgctggc caagcatgtc atcactctgc acgtgagcgc
     1741 actgacacag acacaggctg tggagggcga gattgacctg gccaagctga agaagtttat
     1801 tgcctactgc cgagtgaaat gtggcccccg gctgtcagca gaggctgcag agaaactgaa
     1861 gaaccgctac atcatcatgc ggagcggggc ccgtcagcac gagagggaca gtgaccgccg
     1921 ctccagcatc cccatcactg tgcggcagct ggaggccatt gtgcgcatcg cggaagccct
     1981 cagcaagatg aagctgcagc ctttcgccac agaggcagat gtggaggagg ccctgcggct
     2041 cttccaagtg tccacgttgg atgctgcctt gtccggtacc ctgtcagggg tggagggctt
     2101 caccagccag gaggaccagg agatgctgag ccgcatcgag aagcagctca agcgccgctt
     2161 tgccattggc tcccaggtgt ctgagcacag catcatcaag gacttcacca agcagaaata
     2221 cccggagcac gccatccaca aggtgctgca gctcatgctg cggcgcggcg agatccagca
     2281 tcgcatgcag cgcaaggttc tctaccgcct caagtgacgg ccgcccgcgt tgatctcgat
     2341 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     2401 gaaataattt tgtttaactt taagaaggag atataccatg caagcgggcc cggccttcat
     2461 ggcgggtacc gtggtgctgg acgatgtgga gctgcgggag gctcagagag attacctgga
     2521 cttcctggac gacgaggaag accagggaat ttatcagagc aaagttcggg agctgatcag
     2581 tgacaaccaa taccggctga ttgtcaatgt gaatgacctg cgcaggaaaa acgagaagag
     2641 ggctaaccgg cttctgaaca atgcctttga ggagctggtt gccttccagc gggccttaaa
     2701 ggattttgtg gcctccattg atgctaccta tgccaagcag tatgaggagt tctacgtagg
     2761 actggaaggc agctttggct ccaagcacgt ctccccgcgg actcttacct cctgcttcct
     2821 cagctgtgtg gtctgtgtgg agggcattgt cactaaatgt tctctagttc gtcccaaagt
     2881 cgtccgcagt gtccactact gtcctgctac taagaagacc atagagcgac gttattctga
     2941 tctcaccacc ctggtggcct ttccctccag ctctgtctat cctaccaagg atgaggagaa
     3001 caatcccctt gagacagaat atggcctttc tgtctacaag gatcaccaga ccatcaccat
     3061 ccaggagatg ccggagaagg ccccagccgg ccagctcccc cgctctgtgg acgtcattct
     3121 ggatgatgac ttggtggata aagcgaagcc tggtgaccgg gttcaggtgg tgggaaccta
     3181 ccgttgcctt cctggaaaga agggaggcta cacctctggg accttcagga ctgtcctgat
     3241 tgcctgtaat gttaagcaga tgagcaagga tgctcagccc tctttctctg ctgaggatat
     3301 agccaagatc aagaagttca gtaaaacccg atccaaggat atctttgacc agctggccaa
     3361 gtcattggcc ccaagtatcc atgggcatga ctatgtcaag aaagcaatcc tctgcttgct
     3421 cttgggaggg gtggaacgag acctagaaaa tggcagccac atccgtgggg acatcaatat
     3481 tcttctaata ggagacccat ccgttgccaa gtctcagctt ctgcggtatg tgctttgcac
     3541 tgcaccccga gctatcccca ccactggccg gggctcctct ggagtgggtc tgacggctgc
     3601 tgtcaccaca gaccaggaaa caggagagcg ccgtctggaa gcaggggcca tggtcctggc
     3661 tgaccgaggc gtggtttgca ttgatgaatt tgacaaaatg tctgacatgg atcgcacagc
     3721 catccatgaa gtgatggagc agggtcgagt gaccattgcc aaggctggca tccatgctcg
     3781 gctgaatgcc cgctgcagtg ttttggcagc tgccaaccct gtctacggca ggtatgacca
     3841 gtataagact ccaatggaga acattgggct acaggactca ctgctgtcac gatttgactt
     3901 gctcttcatc atgctggatc agatggatcc tgagcaggat cgggagatct cagaccatgt
     3961 ccttcggatg caccgttaca gagcacctgg ggagcaggat ggcgatgcta tgcccttggg
     4021 tagtgctgtg gatatcctgg ccacagatga tcccaacttt agccaggaag atcagcagga
     4081 cacccagatt tatgagaagc atgacaacct tctacatggg accaagaaga aaaaggagaa
     4141 gatggtgagt gcagcattca tgaagaagta catccatgtg gccaaaatca tcaagcctgt
     4201 cctgacacag gagtcggcca cctacattgc agaagagtat tcacgcctgc gcagccagga
     4261 tagcatgagc tcagacaccg ccaggacatc tccagttaca gcccgaacac tggaaactct
     4321 gattcgactg gccacagccc atgcgaaggc ccgcatgagc aagactgtgg acctgcagga
     4381 tgcagaggaa gctgtggagt tggtccagta tgcttacttt aagaaggttc tggagaagga
     4441 gaagaaacgt aagaagcgaa gtgaggatga atcagagaca gaagatgaag aggagaaaag
     4501 ccaagaggac caggagcaga agaggaagag aaggaagact cgccagccag atgccaaaga
     4561 tggggattca tacgacccct atgacttcag tgacacagag gaggaaatgc ctcaagtaca
     4621 cactccaaag acggcagact cacaggagac caaggaatcc cagaaagtgg agttgagtga
     4681 atccaggttg aaggcattca aggtggccct cttggatgtg ttccgggaag ctcatgcgca
     4741 gtcaatcggc atgaatcgcc tcacagaatc catcaaccgg gacagcgaag agcccttctc
     4801 ttcagttgag atccaggctg ctctgagcaa gatgcaggat gacaatcagg tcatggtgtc
     4861 tgagggcatc atcttcctca tctgaccggg cttctcctca aatctcgagt ctggtaaaga
     4921 aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt
     4981 aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     5041 ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct
     5101 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
     5161 ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt cggggaaatg
     5221 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5281 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     5341 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     5401 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     5461 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5521 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     5581 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     5641 cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca
     5701 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     5761 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     5821 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     5881 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     5941 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     6001 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     6061 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa
     6121 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6181 tagaaaaata aacaaatagg catgcagcgc tcttccgctt cctcgctcac tgactcgcta
     6241 cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact caaaagcggt aatacggtta
     6301 tccacagaat caggggataa agccggaaag aacatgtgag caaaaagcaa agcaccggaa
     6361 gaagccaacg ccgcaggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6421 tcgacgctca agccagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6481 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6541 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag
     6601 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6661 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6721 gccactggca gcagccattg gtaactgatt tagaggactt tgtcttgaag ttatgcacct
     6781 gttaaggcta aactgaaaga acagattttg gtgagtgcgg tcctccaacc cacttacctt
     6841 ggttcaaaga gttggtagct cagcgaacct tgagaaaacc accgttggta gcggtggttt
     6901 ttctttattt atgagatgat gaatcaatcg gtctatcaag tcaacgaaca gctattccgt
     6961 tactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac
     7021 gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg
     7081 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     7141 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7201 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     7261 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     7321 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     7381 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     7441 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     7501 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     7561 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     7621 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     7681 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     7741 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     7801 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     7861 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     7921 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     7981 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     8041 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     8101 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     8161 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     8221 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     8281 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     8341 tgcgactcct gcattaggaa attaatacga ctcactata
//
