LOCUS       sequence-197467-        9195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of S. cerevisiea RPA in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9195)
  AUTHORS   Sibenaller ZA, Sorensen BR, Wold MS
  TITLE     The 32- and 14-kilodalton subunits of replication protein A are 
            responsible for species-specific interactions with single-stranded 
            DNA.
  JOURNAL   Biochemistry. 1998 Sep 8;37(36):12496-506.
  PUBMED    9730822
REFERENCE   2  (bases 1 to 9195)
  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: 
            "Biochemistry."; date: "1998-09-8"; volume: "37(36)"; pages: 
            "12496-506"
COMMENT     Sequence Label: p11d-sctRPA
FEATURES             Location/Qualifiers
     source          1..9195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..27)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        95..199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(200..268,269..1060)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGASKGGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(418..437)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1231..1819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1720..1739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1973..1990
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2005..2144
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2070..2089)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2230..2252
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2246..2437)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4792..4811
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4831..4908)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5114..5133
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5217..5235
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5236..5260
                     /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             5275..5297
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        7191..7209
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7210..7234
                     /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             7249..7271
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             8242..8264
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(8913..8931)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8927..8974
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(9149..9154,9155..9171,9172..9177))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
       61 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      121 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      181 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      241 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      301 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      361 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      421 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
      481 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      541 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      601 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      661 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      721 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
      781 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      841 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      901 agcctccaag ggtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      961 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1021 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1081 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1141 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1201 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1261 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1321 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     1381 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1441 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1501 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1561 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1621 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1681 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1741 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1801 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1861 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1921 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1981 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2041 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2101 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     2161 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2221 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2281 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     2341 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     2401 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     2461 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     2521 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     2581 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     2641 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     2701 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     2761 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     2821 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     2881 ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc
     2941 ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca
     3001 cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt
     3061 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg
     3121 gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc
     3181 cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag
     3241 agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat
     3301 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc
     3361 catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc
     3421 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     3481 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     3541 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     3601 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     3661 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     3721 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaactgtcg
     3781 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3841 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     3901 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     3961 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     4021 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4081 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4141 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4201 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4261 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4321 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4381 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4441 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4501 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     4561 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     4621 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     4681 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     4741 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     4801 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     4861 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     4921 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     4981 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     5041 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     5101 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     5161 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     5221 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     5281 taactttaag aaggagatat accatgggca gtgttcaact ttcgaggggc gattttcata
     5341 gcatcttcac caataagcaa aggtacgata atcccaccgg tggcgtttat caagtttata
     5401 acaccaggaa atctgatggg gctaacagca acagaaagaa tttgatcatg atttccgatg
     5461 gtatttacca tatgaaggct ctgttgagaa accaagctgc atccaagttc cagtcaatgg
     5521 aactacaaag gggtgatatc attcgcgtga taattgcaga acctgctatt gtcagggaaa
     5581 gaaagaaata cgttctttta gtagatgact ttgagttggt ccagtcgcgt gctgatatgg
     5641 tcaaccaaac tagtactttt ttggataact atttctcaga gcatccaaat gaaaccttaa
     5701 aagacgaaga tataactgac agtggtaatg ttgccaatca aacaaacgcc agcaatgctg
     5761 gtgtccctga tatgctgcat tcaaactcaa acttgaatgc aaatgagaga aaattcgcca
     5821 atgaaaaccc taattcgcaa aaaaccagac caatttttgc catcgaacaa ctgtctccat
     5881 accaaaacgt ttggactatc aaagcaagag tttcctacaa gggagaaatt aaaacgtggc
     5941 acaatcaaag aggtgatggt aaactattca atgtcaactt cttggatacc tctggagaaa
     6001 tccgagccac ggcgtttaat gattttgcta caaaatttaa cgaaatttta caagaaggca
     6061 aagtatacta cgtatcaaag gcaaaactcc aaccagctaa gccccaattt actaatctaa
     6121 cacaccctta tgaactgaat ttggatagag acactgttat agaagaatgt ttcgatgaaa
     6181 gtaatgttcc gaaaacccat ttcaatttca tcaaactaga tgctattcag aaccaggaag
     6241 taaattccaa cgtagacgtc ctcggtatta tccaaactat aaacccacat tttgagctaa
     6301 cttcaagggc tgggaagaaa ttcgatcgtc gtgacatcac aattgttgac gactctgggt
     6361 tttctatctc tgttggccta tggaatcagc aagcccttga tttcaacctt cctgaaggtt
     6421 ctgttgctgc cattaaaggt gttcgtgtga cggattttgg tggcaaatct ttgtctatgg
     6481 gattttctag taccctgatt ccgaatccag aaattcctga ggcatatgcc ttaaagggtt
     6541 ggtatgattc caagggccgc aacgcaaact tcatcacttt aaagcaagaa cccggtatgg
     6601 gtggtcaatc ggctgctagc ttaacaaaat tcattgctca gcgtattact attgctagag
     6661 ctcaagctga gaatctagga agaagcgaga aaggtgactt ttttagtgtt aaagctgcta
     6721 taagtttctt aaaagttgat aattttgcat atcctgcctg ttctaatgag aattgtaata
     6781 agaaagttct ggaacagcct gatggtactt ggagatgtga gaagtgcgac accaataatg
     6841 caaggccaaa ttggagatac atcttgacaa tatcaattat tgacgaaacc aatcaactat
     6901 ggctcacttt atttgacgac caagctaaac aattattagg tgttgatgct aatacattaa
     6961 tgtctttgaa ggaagaagac cccaacgaat tcacaaaaat tactcaaagt atccaaatga
     7021 acgaatatga ctttaggatt agagcgcgtg aggatacata caatgatcaa agcagaatta
     7081 gatataccgt tgctaaccta cacagcttga attacagggc tgaagccgac tatcttgccg
     7141 atgagttatc caaggctttg ttagcttaat ggatctcgat cccgcgaaat taatacgact
     7201 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     7261 taagaaggag atatacatat ggcaagttat caaccatata acgaatattc atcagtaacg
     7321 ggcggtggct ttgagaactc tgagtcccgc ccaggtagtg gggagtcgga aactaacact
     7381 agagttaaca ccttgacacc tgtgacgatc aaacaaattc tagagtccaa acaggatatt
     7441 caggacggcc ccttcgtttc gcataaccaa gaacttcatc acgtttgttt tgtaggtgtg
     7501 gtgagaaaca ttacagacca tactgcaaat atttttttaa ctattgagga tggaactggt
     7561 caaatagaag tgagaaaatg gagcgaagat gcaaatgact tggctgccgg taacgatgac
     7621 tcttctggta aaggttatgg ttcgcaagtc gcccaacaat ttgaaattgg cggttacgta
     7681 aaagtttttg gtgctttgaa agagtttggt ggtaagaaaa atatacagta tgcggtgatt
     7741 aagcccatag attcattcaa tgaagtgttg acgcatcact tggaagtcat caaatgtcat
     7801 tccatagcca gtggaatgat gaaacaacct ttggagagtg catccaacaa caatgggcaa
     7861 tcattatttg tcaaggatga taacgataca tcttccggct ccagtccgtt acaaagaatt
     7921 ctagaatttt gtaagaagca atgtgagggc aaagacgcta attcattcgc tgttcccatt
     7981 ccattgatct cgcaatcctt gaatttggat gaaactaccg tcagaaactg ctgtacgacc
     8041 ttgactgacc agggttttat ctacccaact tttgatgaca ataacttctt tgccctatga
     8101 aaatatatag attatttttt tagcatttct tagtattcgt ttgtgtattt ctactaggta
     8161 agtaccaaaa tttgccggaa atagagtgga aaatggaacc atcgaattcc tgcagcccgg
     8221 gggatcccct ctagaaataa ttttgtttaa ctttaagaag gagatgatcc atggccagcg
     8281 aaacaccaag agttgacccc acagaaatct ccaacgtcaa tgctcctgtg tttaggataa
     8341 tagcgcaaat caaatcacaa cccactgaat ctcagttgat tttacaatcg ccaaccatat
     8401 catcgaaaaa cggcagcgag gttgaaatga ttacgttgaa caatattcgt gtatctatga
     8461 ataaaacatt tgagatcgac tcgtggtatg agtttgtttg caggaacaat gatgacggcg
     8521 agctaggatt tttgattcta gacgctgtac tatgcaaatt caaggagaac gaagatttaa
     8581 gcttaaacgg tgtggttgct ttacagagac tatgtaagaa atacccagaa atatactagc
     8641 taaaatcgct tttattctta cctttatgta ctgttgcctc tatatgtctt cccttcatat
     8701 aataaattcc atttctaaaa aaattgtagt tgtgtttata gatgtataga tagataaatg
     8761 aatgtgtact tgctgcctta acgccttccg ctaaaaactt tttgatattt attttatttt
     8821 taacctttcc tgacttcaac ccttactgta atgatgatag taggatccgg ctgctaacaa
     8881 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     8941 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     9001 tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg
     9061 acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt
     9121 tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca
     9181 tgagaattct tgaag
//
