LOCUS       sequence-413704-        8267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human Replication Protein A 70 and 14-kDa subunits in 
            E. coli with 14 having a His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8267)
  AUTHORS   Henricksen LA, Umbricht CB, Wold MS
  TITLE     Recombinant replication protein A: expression, complex formation, 
            and functional characterization.
  JOURNAL   J Biol Chem. 1994 Apr 15;269(15):11121-32.
  PUBMED    8157639
REFERENCE   2  (bases 1 to 8267)
  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: "J Biol 
            Chem."; date: "1994-04-15"; volume: "269(15)"; pages: "11121-32"
COMMENT     Sequence Label: p11d-RPA70/14 (MSW#134)
FEATURES             Location/Qualifiers
     source          1..8267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1110..1129
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1149..1226)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1432..1451
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1535..1554
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1535..1553
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1554..1578
                     /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             1593..1615
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             3981..4003
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        4680..4784
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4785..4853,4854..5645)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5003..5022)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5816..6404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6305..6324
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6558..6575
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6590..6729
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6655..6674)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6815..6837
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6831..7022)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
       61 ttgcgctcac tgcccgcttt ccagtcggga aactgtcgtg ccagctgcat taatgaatcg
      121 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
      181 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
      241 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
      301 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
      361 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
      421 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
      481 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
      541 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
      601 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
      661 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
      721 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
      781 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
      841 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
      901 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
      961 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     1021 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     1081 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     1141 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     1201 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     1261 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     1321 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     1381 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     1441 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     1501 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg
     1561 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     1621 catggtcggc cagctgagcg agggggccat tgcggccatc atgcagaagg gggatacaaa
     1681 cataaagccc atcctccaag tcatcaacat ccgtcccatt actacgggga atagtccgcc
     1741 gcgttatcga ctgctcatga gtgatggatt gaacactcta tcctctttca tgttggcgac
     1801 acagttgaac cctctcgtgg aggaagaaca attgtccagc aactgtgtat gccagattca
     1861 cagatttatt gtgaacactc tgaaagacgg aaggagagta gttatcttga tggaattaga
     1921 agttttgaag tcagctgaag cagttggagt gaagattggc aatccagtgc cctataatga
     1981 aggactcggg cagccgcaag tagctcctcc agcgccagca gccagcccag cagcaagcag
     2041 caggccccag ccgcagaatg gaagctcggg aatgggttct actgtttcta aggcttatgg
     2101 tgcttcaaag acatttggaa aagctgcagg tcccagcctg tcacacactt ctgggggaac
     2161 acagtccaaa gtggtgccca ttgccagcct cactccttac cagtccaagt ggaccatttg
     2221 tgctcgtgtt accaacaaaa gtcagatccg tacctggagc aactcccgag gggaagggaa
     2281 gcttttctcc ctagaactgg ttgacgaaag tggtgaaatc cgagctacag ctttcaatga
     2341 gcaagtggac aagttctttc ctcttattga agtgaacaag gtgtattatt tctcgaaagg
     2401 caccctgaag attgctaaca agcagttcac agctgttaaa aatgactacg agatgacctt
     2461 caataacgag acttccgtca tgccctgtga ggacgaccat catttaccta cggttcagtt
     2521 tgatttcacg gggattgatg acctcgagaa caagtcgaaa gactcacttg tagacatcat
     2581 cgggatctgc aagagctatg aagacgccac taaaatcaca gtgaggtcta acaacagaga
     2641 agttgccaag aggaatatct acttgatgga cacatccggg aaggtggtga ctgctacact
     2701 gtggggggaa gatgctgata aatttgatgg ttctagacag cccgtgttgg ctatcaaagg
     2761 agcccgagtc tctgatttcg gtggacggag cctctccgtg ctgtcttcaa gcactatcat
     2821 tgcgaatcct gacatcccag aggcctataa gcttcgtgga tggtttgacg cagaaggaca
     2881 agccttagat ggtgtttcca tctctgatct aaagagcggc ggagtcggag ggagtaacac
     2941 caactggaaa accttgtatg aggtcaaatc cgagaacctg ggccaaggcg acaagccgga
     3001 ctactttagt tctgtggcca cagtggtgta tcttcgcaaa gagaactgca tgtaccaagc
     3061 ctgcccgact caggactgca ataagaaagt gattgatcaa cagaatggat tgtaccgctg
     3121 tgagaagtgc gacaccgaat ttcccaattt caagtaccgc atgatcctgt cagtaaatat
     3181 tgcagatttt caagagaatc agtgggtgac ttgtttccag gagtctgctg aagctatcct
     3241 tggacaaaat gctgcttatc ttggggaatt aaaagacaag aatgaacagg catttgaaga
     3301 agttttccag aatgccaact tccgatcttt catattcaga gtcagggtca aagtggagac
     3361 ctacaacgac gagtctcgaa ttaaggccac tgtgatggac gtgaagcccg tggactacag
     3421 agagtatggc cgaaggctgg tcatgagcat caggagaagt gcattgatgt gagaggagca
     3481 gtgccaatcg ggcagaagtt tgcaaatagg cagaatggaa tcgatttcct cccacctccg
     3541 tgtgacgatc ccatgttagc tacacagtgc agaggctctt gatggtggac taagcaattt
     3601 cctccctcgt gcgcatctca gaacccatcg gtaggcaaag gaaaatacgc tcaggtggtt
     3661 gtggtgtaga ctgtgtcagg cctacggagt cagccagtgg ctagcgcaag accagtcact
     3721 ccctctgcct tcaggcttct gtcaatttca ttatcatcaa gcaggaatta tgtcgtaagt
     3781 cactgaccct aactgcagac catgaagtaa attatgtaac taggtttttg cttctccagt
     3841 ggtgaccacc cccccccatc cccgctcaca acttgggttc ttctcagcgg ggcgagctga
     3901 gaagcggtca tgagcacctg gggattttag taagtgtgtc ttcctagaat tcctgcagcc
     3961 cgggggatcc tagaaataat tttgtttaac tttaagaagg agatatacca tggtggacat
     4021 gatggacttg cccaggtcgc gcatcaacgc cggcatgcta gctcaattca tcgacaagcc
     4081 tgtctgcttc gtagggaggc tggaaaagat tcatcccacc ggaaaaatgt ttattctttc
     4141 agatggagaa ggaaaaaatg gaaccatcga gttgatggaa ccccttgatg aagaaatctc
     4201 tggaattgtg gaagtggttg gaagagtaac cgccaaggcc accatcttgt gtacatctta
     4261 tgtccagttt aaagaagata gccatccttt tgatcttgga ctttacaatg aagctgtgaa
     4321 aattatccat gacttccctc agttttatcc tttagggatt gtgcaacatg attgatcttg
     4381 atggattttc atacgattgt aaatgagcta tattaaagtc tattaaagga agcccttctt
     4441 gtttgaggga gagatttctg tgctttctca tatttaattt gctgttttta agatattcca
     4501 acctagagtt tttgatggaa ctgatatatt gacagttctc accgaagtcc ttttataaag
     4561 aattgctact ccaatatatg gtcagattag atgcaagaat aaagcagttg tccgagtcta
     4621 agtttctatt ttattaataa aaactaaaat ggtacggtgg cacttttcgg ggaaatgtgc
     4681 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4741 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4801 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4861 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4921 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4981 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     5041 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5101 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5161 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5221 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5281 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     5341 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5401 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5461 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5521 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5581 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5641 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5701 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5761 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5821 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5881 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5941 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6001 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6061 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6121 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6181 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6241 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6301 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6361 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6421 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6481 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6541 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     6601 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     6661 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     6721 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6781 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6841 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     6901 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     6961 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     7021 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     7081 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     7141 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     7201 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     7261 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     7321 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     7381 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     7441 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca
     7501 acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat ggatatgttc
     7561 tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc tccaattctt
     7621 ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc
     7681 tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc
     7741 atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc
     7801 caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt
     7861 cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag
     7921 cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt
     7981 agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     8041 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     8101 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     8161 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     8221 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggct
//
