LOCUS       sequence-197346-        9777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human RPA in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9777)
  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 9777)
  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-tRPA(123)
FEATURES             Location/Qualifiers
     source          1..9777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..21)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             74..96
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        773..877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(878..946,947..1738)
                     /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(1096..1115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1909..2497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2398..2417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2651..2668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2683..2822
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2748..2767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2908..2930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2924..3115)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5470..5489
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5509..5586)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5792..5811
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5895..5914
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5895..5913
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5914..5938
                     /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             5953..5975
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             8341..8363
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(9298..9316)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      9312..9359
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(9725..9730,9731..9747,9748..9753))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga
       61 tcctagaaat aattttgttt aactttaaga aggagatata ccatggtgga catgatggac
      121 ttgcccaggt cgcgcatcaa cgccggcatg ctagctcaat tcatcgacaa gcctgtctgc
      181 ttcgtaggga ggctggaaaa gattcatccc accggaaaaa tgtttattct ttcagatgga
      241 gaaggaaaaa atggaaccat cgagttgatg gaaccccttg atgaagaaat ctctggaatt
      301 gtggaagtgg ttggaagagt aaccgccaag gccaccatct tgtgtacatc ttatgtccag
      361 tttaaagaag atagccatcc ttttgatctt ggactttaca atgaagctgt gaaaattatc
      421 catgacttcc ctcagtttta tcctttaggg attgtgcaac atgattgatc ttgatggatt
      481 ttcatacgat tgtaaatgag ctatattaaa gtctattaaa ggaagccctt cttgtttgag
      541 ggagagattt ctgtgctttc tcatatttaa tttgctgttt ttaagatatt ccaacctaga
      601 gtttttgatg gaactgatat attgacagtt ctcaccgaag tccttttata aagaattgct
      661 actccaatat atggtcagat tagatgcaag aataaagcag ttgtccgagt ctaagtttct
      721 attttattaa taaaaactaa aatggtacgg tggcactttt cggggaaatg tgcgcggaac
      781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      841 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      901 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      961 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1021 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1081 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     1141 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1201 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1261 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1321 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1381 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1441 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1501 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1561 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1621 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1681 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1741 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1801 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1861 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1921 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1981 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2041 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2101 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2161 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2221 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2281 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2341 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2401 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2461 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2521 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2581 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2641 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2701 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2761 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2821 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2881 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2941 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3001 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3061 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3121 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3181 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3241 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3301 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3361 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3421 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3481 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3541 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     3601 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     3661 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     3721 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     3781 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     3841 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
     3901 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     3961 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     4021 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     4081 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     4141 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     4201 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     4261 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     4321 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     4381 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     4441 ccagtcggga aactgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4501 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4561 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4621 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4681 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     4741 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4801 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     4861 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     4921 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     4981 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     5041 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5101 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     5161 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5221 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5281 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5341 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     5401 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     5461 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     5521 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5581 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     5641 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     5701 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     5761 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     5821 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     5881 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     5941 tctagaaata attttgttta actttaagaa ggagatatac catggtcggc cagctgagcg
     6001 agggggccat tgcggccatc atgcagaagg gggatacaaa cataaagccc atcctccaag
     6061 tcatcaacat ccgtcccatt actacgggga atagtccgcc gcgttatcga ctgctcatga
     6121 gtgatggatt gaacactcta tcctctttca tgttggcgac acagttgaac cctctcgtgg
     6181 aggaagaaca attgtccagc aactgtgtat gccagattca cagatttatt gtgaacactc
     6241 tgaaagacgg aaggagagta gttatcttga tggaattaga agttttgaag tcagctgaag
     6301 cagttggagt gaagattggc aatccagtgc cctataatga aggactcggg cagccgcaag
     6361 tagctcctcc agcgccagca gccagcccag cagcaagcag caggccccag ccgcagaatg
     6421 gaagctcggg aatgggttct actgtttcta aggcttatgg tgcttcaaag acatttggaa
     6481 aagctgcagg tcccagcctg tcacacactt ctgggggaac acagtccaaa gtggtgccca
     6541 ttgccagcct cactccttac cagtccaagt ggaccatttg tgctcgtgtt accaacaaaa
     6601 gtcagatccg tacctggagc aactcccgag gggaagggaa gcttttctcc ctagaactgg
     6661 ttgacgaaag tggtgaaatc cgagctacag ctttcaatga gcaagtggac aagttctttc
     6721 ctcttattga agtgaacaag gtgtattatt tctcgaaagg caccctgaag attgctaaca
     6781 agcagttcac agctgttaaa aatgactacg agatgacctt caataacgag acttccgtca
     6841 tgccctgtga ggacgaccat catttaccta cggttcagtt tgatttcacg gggattgatg
     6901 acctcgagaa caagtcgaaa gactcacttg tagacatcat cgggatctgc aagagctatg
     6961 aagacgccac taaaatcaca gtgaggtcta acaacagaga agttgccaag aggaatatct
     7021 acttgatgga cacatccggg aaggtggtga ctgctacact gtggggggaa gatgctgata
     7081 aatttgatgg ttctagacag cccgtgttgg ctatcaaagg agcccgagtc tctgatttcg
     7141 gtggacggag cctctccgtg ctgtcttcaa gcactatcat tgcgaatcct gacatcccag
     7201 aggcctataa gcttcgtgga tggtttgacg cagaaggaca agccttagat ggtgtttcca
     7261 tctctgatct aaagagcggc ggagtcggag ggagtaacac caactggaaa accttgtatg
     7321 aggtcaaatc cgagaacctg ggccaaggcg acaagccgga ctactttagt tctgtggcca
     7381 cagtggtgta tcttcgcaaa gagaactgca tgtaccaagc ctgcccgact caggactgca
     7441 ataagaaagt gattgatcaa cagaatggat tgtaccgctg tgagaagtgc gacaccgaat
     7501 ttcccaattt caagtaccgc atgatcctgt cagtaaatat tgcagatttt caagagaatc
     7561 agtgggtgac ttgtttccag gagtctgctg aagctatcct tggacaaaat gctgcttatc
     7621 ttggggaatt aaaagacaag aatgaacagg catttgaaga agttttccag aatgccaact
     7681 tccgatcttt catattcaga gtcagggtca aagtggagac ctacaacgac gagtctcgaa
     7741 ttaaggccac tgtgatggac gtgaagcccg tggactacag agagtatggc cgaaggctgg
     7801 tcatgagcat caggagaagt gcattgatgt gagaggagca gtgccaatcg ggcagaagtt
     7861 tgcaaatagg cagaatggaa tcgatttcct cccacctccg tgtgacgatc ccatgttagc
     7921 tacacagtgc agaggctctt gatggtggac taagcaattt cctccctcgt gcgcatctca
     7981 gaacccatcg gtaggcaaag gaaaatacgc tcaggtggtt gtggtgtaga ctgtgtcagg
     8041 cctacggagt cagccagtgg ctagcgcaag accagtcact ccctctgcct tcaggcttct
     8101 gtcaatttca ttatcatcaa gcaggaatta tgtcgtaagt cactgaccct aactgcagac
     8161 catgaagtaa attatgtaac taggtttttg cttctccagt ggtgaccacc cccccccatc
     8221 cccgctcaca acttgggttc ttctcagcgg ggcgagctga gaagcggtca tgagcacctg
     8281 gggattttag taagtgtgtc ttcctagaat tcctgcagcc cgggggatcc tagaaataat
     8341 tttgtttaac tttaagaagg agatatacat atgtggaaca gtggattcga aagctatggc
     8401 agctcctcat acgggggagc cggcggctac acgcagtccc cggggggctt tggatcgccc
     8461 gcaccttctc aagccgaaaa gaaatcaaga gcccgagccc agcacattgt gccctgtact
     8521 atatctcagc tgctttctgc cactttggtt gatgaagtgt tcagaattgg gaatgttgag
     8581 atttcacagg tcactattgt ggggatcatc agacatgcag agaaggctcc aaccaacatt
     8641 gtttacaaaa tagatgacat gacagctgca cccatggacg ttcgccagtg ggttgacaca
     8701 gatgacacca gcagtgaaaa cactgtggtt cctccagaaa catatgtgaa agtggcaggc
     8761 cacctgagat cttttcagaa caaaaagagc ctggtagcct ttaagatcat gcccctggag
     8821 gatatgaatg agttcaccac acatattctg gaagtgatca atgcacacat ggtactaagc
     8881 aaagccaaca gccagccctc agcagggaga gcacctatca gcaatccagg aatgagtgaa
     8941 gcagggaact ttggtgggaa tagcttcatg ccagcaaatg gcctcactgt ggcccaaaac
     9001 caggtgttga atttgattaa ggcttgtcca agacctgaag ggttgaactt tcaggatctc
     9061 aagaaccagc tgaaacacat gtctgtatcc tcaatcaagc aagctgtgga ttttctgagc
     9121 aatgaggggc acatctattc tactgtggat gatgaccatt ttaaatccac agatgcagaa
     9181 taactggatc taactgggta cctgagatat tttacagctg gacctagttt cacaatctgt
     9241 tgtctccgga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     9301 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     9361 tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag
     9421 tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac
     9481 agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg
     9541 ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc
     9601 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     9661 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     9721 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaa
//
