LOCUS       sequence-197485-        9822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human RPA with aspartic acid substituted 
            phosphorylation domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9822)
  AUTHORS   Binz SK, Lao Y, Lowry DF, Wold MS
  TITLE     The phosphorylation domain of the 32-kDa subunit of replication 
            protein A (RPA) modulates RPA-DNA interactions. Evidence for an 
            intersubunit interaction.
  JOURNAL   J Biol Chem. 2003 Sep 12;278(37):35584-91. Epub 2003 Jun 20.
  PUBMED    12819197
REFERENCE   2  (bases 1 to 9822)
  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: "2003-09-12"; volume: "278(37)"; pages: "35584-91. 
            Epub 2003 Jun 20"
COMMENT     Sequence Label: p11d-tRPA-32Asp8
FEATURES             Location/Qualifiers
     source          1..9822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     498..517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     751..768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    783..922
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(848..867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1008..1030
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1024..1215)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3570..3589
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3609..3686)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3892..3911
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3995..4014
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3995..4013
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4014..4038
                     /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             4053..4075
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             6441..6463
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             7170..7192
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(8127..8145)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8141..8188
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(8554..8559,8560..8576,8577..8582))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(8609..8627)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8695..8799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8800..8868,8869..9660)
                     /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(9018..9037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
       61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      121 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
      181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      781 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
      841 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
      901 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      961 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     1021 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     1081 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     1141 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     1201 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     1261 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     1321 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     1381 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     1441 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     1501 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     1561 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     1621 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     1681 tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc
     1741 gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt
     1801 ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc
     1861 gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg
     1921 gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg
     1981 acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc
     2041 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
     2101 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
     2161 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     2221 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     2281 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     2341 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     2401 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     2461 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     2521 ttgcgctcac tgcccgcttt ccagtcggga aactgtcgtg ccagctgcat taatgaatcg
     2581 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     2641 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     2701 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     2761 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     2821 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     2881 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     2941 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     3001 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     3061 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     3121 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     3181 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     3241 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     3301 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     3361 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     3421 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     3481 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     3541 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     3601 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     3661 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     3721 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     3781 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     3841 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     3901 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     3961 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg
     4021 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     4081 catggtcggc cagctgagcg agggggccat tgcggccatc atgcagaagg gggatacaaa
     4141 cataaagccc atcctccaag tcatcaacat ccgtcccatt actacgggga atagtccgcc
     4201 gcgttatcga ctgctcatga gtgatggatt gaacactcta tcctctttca tgttggcgac
     4261 acagttgaac cctctcgtgg aggaagaaca attgtccagc aactgtgtat gccagattca
     4321 cagatttatt gtgaacactc tgaaagacgg aaggagagta gttatcttga tggaattaga
     4381 agttttgaag tcagctgaag cagttggagt gaagattggc aatccagtgc cctataatga
     4441 aggactcggg cagccgcaag tagctcctcc agcgccagca gccagcccag cagcaagcag
     4501 caggccccag ccgcagaatg gaagctcggg aatgggttct actgtttcta aggcttatgg
     4561 tgcttcaaag acatttggaa aagctgcagg tcccagcctg tcacacactt ctgggggaac
     4621 acagtccaaa gtggtgccca ttgccagcct cactccttac cagtccaagt ggaccatttg
     4681 tgctcgtgtt accaacaaaa gtcagatccg tacctggagc aactcccgag gggaagggaa
     4741 gcttttctcc ctagaactgg ttgacgaaag tggtgaaatc cgagctacag ctttcaatga
     4801 gcaagtggac aagttctttc ctcttattga agtgaacaag gtgtattatt tctcgaaagg
     4861 caccctgaag attgctaaca agcagttcac agctgttaaa aatgactacg agatgacctt
     4921 caataacgag acttccgtca tgccctgtga ggacgaccat catttaccta cggttcagtt
     4981 tgatttcacg gggattgatg acctcgagaa caagtcgaaa gactcacttg tagacatcat
     5041 cgggatctgc aagagctatg aagacgccac taaaatcaca gtgaggtcta acaacagaga
     5101 agttgccaag aggaatatct acttgatgga cacatccggg aaggtggtga ctgctacact
     5161 gtggggggaa gatgctgata aatttgatgg ttctagacag cccgtgttgg ctatcaaagg
     5221 agcccgagtc tctgatttcg gtggacggag cctctccgtg ctgtcttcaa gcactatcat
     5281 tgcgaatcct gacatcccag aggcctataa gcttcgtgga tggtttgacg cagaaggaca
     5341 agccttagat ggtgtttcca tctctgatct aaagagcggc ggagtcggag ggagtaacac
     5401 caactggaaa accttgtatg aggtcaaatc cgagaacctg ggccaaggcg acaagccgga
     5461 ctactttagt tctgtggcca cagtggtgta tcttcgcaaa gagaactgca tgtaccaagc
     5521 ctgcccgact caggactgca ataagaaagt gattgatcaa cagaatggat tgtaccgctg
     5581 tgagaagtgc gacaccgaat ttcccaattt caagtaccgc atgatcctgt cagtaaatat
     5641 tgcagatttt caagagaatc agtgggtgac ttgtttccag gagtctgctg aagctatcct
     5701 tggacaaaat gctgcttatc ttggggaatt aaaagacaag aatgaacagg catttgaaga
     5761 agttttccag aatgccaact tccgatcttt catattcaga gtcagggtca aagtggagac
     5821 ctacaacgac gagtctcgaa ttaaggccac tgtgatggac gtgaagcccg tggactacag
     5881 agagtatggc cgaaggctgg tcatgagcat caggagaagt gcattgatgt gagaggagca
     5941 gtgccaatcg ggcagaagtt tgcaaatagg cagaatggaa tcgatttcct cccacctccg
     6001 tgtgacgatc ccatgttagc tacacagtgc agaggctctt gatggtggac taagcaattt
     6061 cctccctcgt gcgcatctca gaacccatcg gtaggcaaag gaaaatacgc tcaggtggtt
     6121 gtggtgtaga ctgtgtcagg cctacggagt cagccagtgg ctagcgcaag accagtcact
     6181 ccctctgcct tcaggcttct gtcaatttca ttatcatcaa gcaggaatta tgtcgtaagt
     6241 cactgaccct aactgcagac catgaagtaa attatgtaac taggtttttg cttctccagt
     6301 ggtgaccacc cccccccatc cccgctcaca acttgggttc ttctcagcgg ggcgagctga
     6361 gaagcggtca tgagcacctg gggattttag taagtgtgtc ttcctagaat tcctgcagcc
     6421 cgggggatcc tagaaataat tttgtttaac tttaagaagg agatatacca tggtggacat
     6481 gatggacttg cccaggtcgc gcatcaacgc cggcatgcta gctcaattca tcgacaagcc
     6541 tgtctgcttc gtagggaggc tggaaaagat tcatcccacc ggaaaaatgt ttattctttc
     6601 agatggagaa ggaaaaaatg gaaccatcga gttgatggaa ccccttgatg aagaaatctc
     6661 tggaattgtg gaagtggttg gaagagtaac cgccaaggcc accatcttgt gtacatctta
     6721 tgtccagttt aaagaagata gccatccttt tgatcttgga ctttacaatg aagctgtgaa
     6781 aattatccat gacttccctc agttttatcc tttagggatt gtgcaacatg attgatcttg
     6841 atggattttc atacgattgt aaatgagcta tattaaagtc tattaaagga agcccttctt
     6901 gtttgaggga gagatttctg tgctttctca tatttaattt gctgttttta agatattcca
     6961 acctagagtt tttgatggaa ctgatatatt gacagttctc accgaagtcc ttttataaag
     7021 aattgctact ccaatatatg gtcagattag atgcaagaat aaagcagttg tccgagtcta
     7081 agtttctatt ttattaataa aaactaaaat ggtacgtaca aaaaaaaaac cggaattcga
     7141 gctcggtacc cggggatcct agaaataatt ttgtttaact ttaagaagga gatatacata
     7201 tgtggaacag tggattcgaa gactatggcg acgacgacta cgggggagcc ggcggctacg
     7261 accaggatcc ggggggcttt ggagacccag cacctgatca agccgaaaag aaatcaagag
     7321 cccgagccca gcacattgtg ccctgtacta tatctcagct gctttctgcc actttggttg
     7381 atgaagtgtt cagaattggg aatgttgaga tttcacaggt cactattgtg gggatcatca
     7441 gacatgcaga gaaggctcca atcaacattg tttacaaaat agatgacatg acagctgcac
     7501 ccatggacgt tcgccagtgg gttgacacag atgacaccag cagtgaaaac actgtggttc
     7561 ctccagaaac atatgtgaaa gtggcaggcc acctgagatc ttttcagaac aaaaagagcc
     7621 tggtagcctt taagatcatg cccctggagg atatgaatga gttcaccaca catattctgg
     7681 aagtgatcaa tgcacacatg gtactaagca aagccaacag ccagccctca gcagggagag
     7741 cacctatcag caatccagga atgagtgaag cagggaactt tggtgggaat agcttcatgc
     7801 cagcaaatgg cctcactgtg gcccaaaacc aggtgttgaa tttgattaag gcttgtccaa
     7861 gacctgaagg gttgaacttt caggatctca agaaccagct gaaacacatg tctgtatcct
     7921 caatcaagca agctgtggat tttctgagca atgaggggca catctattct actgtggatg
     7981 atgaccattt taaatccaca gatgcagaat aactggatct aactgggtac ctgagatatt
     8041 ttacagctgg acctagtttc acaatctgtt gtctccggat ccggctgcta acaaagcccg
     8101 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     8161 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca
     8221 ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca
     8281 ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt
     8341 gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga
     8401 cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca
     8461 gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac
     8521 tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc
     8581 aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     8641 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     8701 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     8761 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     8821 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     8881 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     8941 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     9001 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     9061 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     9121 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     9181 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     9241 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     9301 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     9361 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     9421 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     9481 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     9541 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     9601 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     9661 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     9721 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     9781 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aa
//
