LOCUS       sequence-227777-        8757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human MCM4 and human MCM6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8757)
  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 8757)
  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: pCDF-MCM4/6
FEATURES             Location/Qualifiers
     source          1..8757
                     /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        2612..2630
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2631..2655
                     /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             2670..2692
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5342..5386
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(5424..5442)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5438..5485
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5656..6447)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(6448..6539)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6587..7325)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(7535..8617)
                     /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     8579..8598
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8618..8695)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(8741..8757,1..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 agatggacct
      121 cgcggcggca gcggagccgg gcgccggcag ccagcacctg gaggtccgcg acgaggtggc
      181 cgagaagtgc cagaaactgt tcctggactt cttggaggag tttcagagca gcgatggaga
      241 aattaaatac ttgcaattag cagaggaact gattcgtcct gagagaaaca cattggttgt
      301 gagttttgtg gacctggaac aatttaacca gcaactttcc accaccattc aagaggagtt
      361 ctatagagtt tacccttacc tgtgtcgggc cttgaaaaca ttcgtcaaag accgtaaaga
      421 gatccctctt gccaaggatt tttatgttgc attccaagac ctgcctacca gacacaagat
      481 tcgagagctc acctcatcca gaattggttt gctcactcgc atcagtgggc aggtggtgcg
      541 gactcaccca gttcacccag agcttgtgag cggaactttt ctgtgcttgg actgtcagac
      601 agtgatcagg gatgtagaac agcagttcaa atacacacag ccaaacatct gccgaaatcc
      661 agtttgtgcc aacaggagga gattcttact ggatacaaat aaatcaagat ttgttgattt
      721 tcaaaaggtt cgtattcaag agacccaagc tgagcttcct cgagggagta tcccccgcag
      781 tttagaagta attttaaggg ctgaagctgt ggaatcagct caagctggtg acaagtgtga
      841 ctttacaggg acactgattg ttgtgcctga cgtctccaag cttagcacac caggagcacg
      901 tgcagaaact aattcccgtg tcagtggtgt tgatggatat gagacagaag gcattcgagg
      961 actccgggcc cttggtgtta gggacctttc ttataggctg gtctttcttg cctgctgtgt
     1021 tgcgccaacc aacccaaggt ttggggggaa agagctcaga gatgaggaac agacagctga
     1081 gagcattaag aaccaaatga ctgtgaaaga atgggagaaa gtgtttgaga tgagtcaaga
     1141 taaaaatcta taccacaatc tttgtaccag cctgttccct actatacatg gcaatgatga
     1201 agtaaaacgg ggtgtcctgc tgatgctctt tggtggcgtt ccaaagacaa caggagaagg
     1261 gacctctctt cgaggggaca taaatgtttg cattgttggt gacccaagta cagctaagag
     1321 ccaatttctc aagcacgtgg aggagttcag ccccagagct gtctacacca gtggtaaagc
     1381 gtccagtgct gctggcttaa cagcagctgt tgtgagagat gaagaatctc atgagtttgt
     1441 cattgaggct ggagctttga tgttggctga taatggtgtg tgttgtattg atgaatttga
     1501 taagatggac gtgcgggatc aagttgctat tcatgaagct atggaacagc agaccatatc
     1561 catcactaaa gcaggagtga aggctactct gaacgcccgg acgtccattt tggcagcagc
     1621 aaacccaatc agtggacact atgacagatc aaaatcattg aaacagaata taaatttgtc
     1681 agctcccatc atgtcccgat tcgatctctt ctttatcctt gtggatgaat gtaatgaggt
     1741 tacagattat gccattgcca ggcgcatagt agatttgcat tcaagaattg aggaatcaat
     1801 tgatcgtgtc tattccctcg atgatatcag aagatatctt ctctttgcaa gacagtttaa
     1861 acccaagatt tccaaagagt cagaggactt cattgtggag caatataaac atctccgcca
     1921 gagagatggt tctggagtga ccaagtcttc atggaggatt acagtgcgac agcttgagag
     1981 catgattcgt ctctctgaag ctatggctcg gatgcactgc tgtgatgagg tccaacctaa
     2041 acatgtgaag gaagctttcc ggttactgaa taaatcaatc atccgtgtgg aaacacctga
     2101 tgtcaatcta gatcaagagg aagagatcca gatggaggta gatgagggtg ctggtggcat
     2161 caatggtcat gctgacagcc ctgctcctgt gaacgggatc aatggctaca atgaagacat
     2221 aaatcaagag tctgctccca aagcctcctt aaggctgggc ttctctgagt actgccgaat
     2281 ctctaacctt attgtgcttc acctcagaaa ggtggaagaa gaagaggacg agtcagcatt
     2341 aaagaggagc gagcttgtta actggtactt gaaggaaatc gaatcagaga tagactctga
     2401 agaagaactt ataaataaaa aaagaatcat agagaaagtt attcatcgac tcacacacta
     2461 tgatcatgtt ctaattgagc tcacccaggc tggattgaaa ggctccacag agggaagtga
     2521 gagctatgaa gaagatccct acttggtagt taaccctaac tacttgctcg aagattgacg
     2581 gccgcccgcg ttgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     2641 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
     2701 gcaagcgggc ccggccttca tgtcgtcccc ggcgtcgacc ccgagccgcc gcggcagccg
     2761 gcgtggaagg gccacccccg cccagacgcc tcggagtgag gatgccaggt catctccctc
     2821 tcagagacgt agaggcgagg attccacctc cacgggggag ttgcagccga tgccaacctc
     2881 gcctggagtg gacctgcaga gccctgctgc gcaggacgtg ctgttttcca gccctcccca
     2941 aatgcattct tcagctatcc ctcttgactt tgatgttagt tcaccactga catacggcac
     3001 tcccagctct cgggtagagg gaaccccaag aagtggtgtt aggggcacac ctgtgagaca
     3061 gaggcctgac ctgggctctg cacagaaggg cctgcaagtg gatctgcagt ctgacggggc
     3121 agcagcagaa gatatagtgg caagtgagca gtctctaggc caaaaacttg tgatctgggg
     3181 aacagatgta aatgtggcag catgcaaaga aaactttcag agatttcttc agcgttttat
     3241 tgaccctctg gctaaagaag aagaaaatgt tggcatagat attactgaac ctctatacat
     3301 gcaacgactt ggggagatta atgttattgg tgagccattt ttaaatgtga actgtgaaca
     3361 catcaaatca tttgacaaaa atttgtacag acaactcatc tcttacccac aggaagttat
     3421 tccaactttt gacatggctg tcaatgaaat cttctttgac cgttaccctg actcaatctt
     3481 agaacatcag attcaagtaa gaccattcaa cgcattgaag actaagaata tgagaaacct
     3541 gaatccagaa gacattgacc agctcatcac catcagcggc atggtgatca ggacatccca
     3601 gctgattccc gagatgcagg aggccttctt ccagtgccaa gtgtgtgccc acacgacccg
     3661 ggtggagatg gaccgcggcc gcattgcaga gcccagtgtg tgcgggcgct gccacaccac
     3721 ccacagcatg gcactcatcc acaaccgctc cctcttctct gacaagcaga tgatcaagct
     3781 tcaggagtct ccggaagaca tgcctgcagg gcagacacca cacacagtta tcctgtttgc
     3841 tcacaatgat ctcgttgaca aggtccagcc tggggacaga gtgaatgtta caggcatcta
     3901 tcgagctgtg cctattcgag tcaatccaag agtgagtaat gtgaagtctg tctacaaaac
     3961 ccacattgat gtcattcatt atcggaaaac ggatgcaaaa cgtctgcatg gccttgatga
     4021 agaagcagaa cagaaacttt tttcagagaa acgtgtggaa ttgcttaagg aactttccag
     4081 gaaaccagac atttatgaga ggcttgcttc agccttggct ccaagcattt atgaacatga
     4141 agatataaag aagggaattt tgcttcagct ctttggcggg acaaggaagg attttagtca
     4201 cactggaagg ggcaaatttc gggctgagat caacatcttg ctgtgtggcg accctggtac
     4261 cagcaagtcc cagctgctgc agtacgtgta caacctcgtc cccaggggcc agtacacgtc
     4321 tgggaagggc tccagtgcag ttggcctcac tgcgtacgta atgaaagacc ctgagacaag
     4381 gcagctggtc ctgcagacag gtgctcttgt cctgagtgac aacggcatct gctgtatcga
     4441 tgagttcgac aagatgaatg aaagtacaag atcggtattg catgaagtca tggaacagca
     4501 gactctgtcc attgcaaagg ctgggatcat ctgtcagctc aatgcgcgca cctctgtcct
     4561 ggcagcagca aatcccattg agtctcagtg gaatcctaaa aaaacaacca ttgaaaacat
     4621 ccagctgcct catactttat tatcaaggtt tgatttgatc ttcctcatgc tggaccctca
     4681 ggacgaagcc tatgacaggc gtctggctca ccacctggtc gcactgtact accagagcga
     4741 ggagcaggca gaggaggagc tcctggacat ggcggtgcta aaggactaca ttgcctacgc
     4801 gcacagcacc atcatgccgc ggctaagtga ggaagccagc caggctctca tcgaggctta
     4861 tgtagacatg aggaagattg gcagtagccg gggaatggtt tctgcatacc ctcgacagct
     4921 agagtcatta atccgcttag cagaagccca tgctaaagta agattgtcta acaaagttga
     4981 agccattgat gtggaagagg ccaaacgcct ccatcgggaa gctctgaagc agtctgcaac
     5041 tgatccccgg actggcatcg tggacatatc tattcttact acggggatga gtgccacctc
     5101 tcgtaaacgg aaagaagaat tagctgaagc attgaaaaag cttattttat ctaagggcaa
     5161 aacaccagct ctaaaatacc agcaactttt tgaagatatt cggggacaat ctgacatagc
     5221 aattactaaa gatatgtttg aagaagcact gcgtgccctg gcagatgatg atttcctgac
     5281 agtgactggg aagaccgtgc gcttgctctg accgggcttc tcctcaaatc tcgagtctgg
     5341 taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc
     5401 ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     5461 taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac
     5521 actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg
     5581 accctgccct gaaccgacga ccgggtcatc gtggccggat cttgcggccc ctcggcttga
     5641 acgaattgtt agacattatt tgccgactac cttggtgatc tcgcctttca cgtagtggac
     5701 aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc
     5761 ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc
     5821 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
     5881 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
     5941 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
     6001 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
     6061 aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc
     6121 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
     6181 ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag
     6241 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
     6301 atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
     6361 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
     6421 tacttcggcg atcaccgctt ccctcatact cttccttttt caatattatt gaagcattta
     6481 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6541 agctagctca ctcggtcgct acgctccggg cgtgagactg cggcgggcgc tgcggacaca
     6601 tacaaagtta cccacagatt ccgtggataa gcaggggact aacatgtgag gcaaaacagc
     6661 agggccgcgc cggtggcgtt tttccatagg ctccgccctc ctgccagagt tcacataaac
     6721 agacgctttt ccggtgcatc tgtgggagcc gtgaggctca accatgaatc tgacagtacg
     6781 ggcgaaaccc gacaggactt aaagatcccc accgtttccg gcgggtcgct ccctcttgcg
     6841 ctctcctgtt ccgaccctgc cgtttaccgg atacctgttc cgcctttctc ccttacggga
     6901 agtgtggcgc tttctcatag ctcacacact ggtatctcgg ctcggtgtag gtcgttcgct
     6961 ccaagctggg ctgtaagcaa gaactccccg ttcagcccga ctgctgcgcc ttatccggta
     7021 actgttcact tgagtccaac ccggaaaagc acggtaaaac gccactggca gcagccattg
     7081 gtaactggga gttcgcagag gatttgttta gctaaacacg cggttgctct tgaagtgtgc
     7141 gccaaagtcc ggctacactg gaaggacaga tttggttgct gtgctctgcg aaagccagtt
     7201 accacggtta agcagttccc caactgactt aaccttcgat caaaccacct ccccaggtgg
     7261 ttttttcgtt tacagggcaa aagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7321 tgatcttttc tactgaaccg ctctagattt cagtgcaatt tatctcttca aatgtagcac
     7381 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
     7441 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     7501 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7561 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7621 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     7681 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     7741 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     7801 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     7861 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     7921 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     7981 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     8041 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     8101 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     8161 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     8221 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     8281 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     8341 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     8401 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     8461 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     8521 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     8581 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     8641 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     8701 tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact cactata
//
