LOCUS       sequence-413079-        7311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human Replication Protein A 4 and His-tagged RPA3 in 
            E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7311)
  AUTHORS   Mason AC, Haring SJ, Pryor JM, Staloch CA, Gan TF, Wold MS
  TITLE     An alternative form of replication protein a prevents viral 
            replication in vitro.
  JOURNAL   J Biol Chem. 2009 Feb 20;284(8):5324-31. doi: 
            10.1074/jbc.M808963200. Epub 2008 Dec 29.
  PUBMED    19116208
REFERENCE   2  (bases 1 to 7311)
  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.M808963200"; journalName: "J Biol Chem"; date: 
            "2009-02-20- 20"; volume: "284"; issue: "8"; pages: "5324-31"
COMMENT     Sequence Label: pET16b-RPA4/His14 (MSW#949)
FEATURES             Location/Qualifiers
     source          1..7311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1964..2155
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2149..2171)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2257..2396
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2312..2331
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2411..2428)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2582..3170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2662..2681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3341..4132,4133..4201))
                     /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     3964..3983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4202..4306)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4337..4383)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     RBS             5180..5202
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(5595..5606)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(5619..5648)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             5661..5683
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    6452..6476
                     /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)."
     primer_bind     complement(6476..6495)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6477..6495)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6579..6598)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6804..6881
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(6901..6920)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
       61 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
      121 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
      181 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
      241 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
      301 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
      361 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
      421 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
      481 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
      541 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
      601 attcattaat gcagctggca cgacagtttc ccgactggaa agcgggcagt gagcgcaacg
      661 caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag
      721 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
      781 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
      841 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
      901 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
      961 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt
     1021 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg
     1081 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc
     1141 ttcaaggatc gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca
     1201 cggcgattta tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg
     1261 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     1321 cctgaatgga agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa
     1381 tcaattcttg cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc
     1441 gtccgccatc tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg
     1501 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     1561 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     1621 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     1681 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     1741 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     1801 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     1861 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     1921 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     1981 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     2041 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     2101 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     2161 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     2221 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     2281 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     2341 caccattgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     2401 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2461 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2521 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2581 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2641 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2701 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2761 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2821 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2881 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2941 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3001 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3061 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3121 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3181 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3241 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3301 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3361 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3421 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3481 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3541 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3601 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     3661 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3721 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3781 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3841 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3901 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     3961 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4021 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4081 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4141 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4201 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4261 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4321 aagtgccacc tgacgtcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     4381 ctattatcaa tcagcagact taaaatgctc ccgatccaca gtgggataga tgtggccctc
     4441 aacggtcaga taatcaatcg cttccttgat ggccttgacg ctaaggtcgc agagctgagc
     4501 ccggagctca tggatgctct tcccttcctg atgaggacac tcatgaatca aacgcagcac
     4561 ttcgtcctgg atgaaattgc ggtgactctc atcgttatcc ccagcatcat tcacttctga
     4621 tggagacaca ggcacacttt ctacagtggt atcacgacgg gctttatcca gcatcatgtg
     4681 tgcattgacc gtttccagaa tatgcacggt gaactcgttc atgtcctcta ggacatgaat
     4741 tttcaatacc tcaaggctct ttgttcccgt gggacatttg aggataccaa acactttgac
     4801 atatactccg actgacaatg gagtcacctg cttgactttc tctctaccaa accactgtcg
     4861 ggcctcgatt ggtttcgcgg tcatatcatc aattttgtaa caaatgtgat ttgaagcctt
     4921 ctctgcccct ctgattaccc ccacgatgga gacctgggaa actataattc ccctaacctt
     4981 gaacacaggg tcaaacacag tagagctgag tagctggttc acgttacacg gtacaacgtc
     5041 ctgaattcgg accttaggtc tttgggtctt aatagcagga gttgcatctc tctcacacag
     5101 ttggtcactg cctccactcg ctccatcagc agcagaaatg ctgccatagc tcccaaaccc
     5161 actcttactc atatgtatat ctccttctta aagttaaaca aaattatttc taggatccat
     5221 caagatcaat catgttgcac aatccctaaa ggataaaact gagggaagtc atggataatt
     5281 ttcacagctt cattgtaaag tccaagatca aaaggatggc tatcttcttt aaactggaca
     5341 taagatgtac acaagatggt ggccttggcg gttactcttc caaccacttc cacaattcca
     5401 gagatttctt catcaagggg ttccatcaac tcgatggttc cattttttcc ttctccatct
     5461 gaaagaataa acatttttcc ggtgggatga atcttttcca gcctccctac gaagcagaca
     5521 ggcttgtcga tgaattgagc tagcatgccg gcgttgatgc gcgacctggg caagtccatc
     5581 atgtccacca tatgacgacc ttcgatatgg ccgctgctgt gatgatgatg atgatgatga
     5641 tgatgatggc ccatggtata tctccttctt aaagttaaac aaaattattt ctaggagatc
     5701 cgctcctctc acatcaatgc acttctcctg atgctcatga ccagccttcg gccatactct
     5761 ctgtagtcca cgggcttcac gtccatcaca gtggccttaa ttcgagactc gtcgttgtag
     5821 gtctccactt tgaccctgac tctgaatatg aaagatcgga agttggcatt ctggaaaact
     5881 tcttcaaatg cctgttcatt cttgtctttt aattccccaa gataagcagc attttgtcca
     5941 aggatagctt cagcagactc ctggaaacaa gtcacccact gattctcttg aaaatctgca
     6001 atatttactg acaggatcat gcggtacttg aaattgggaa attcggtgtc gcacttctca
     6061 cagcggtaca atccattctg ttgatcaatc actttcttat tgcagtcctg agtcgggcag
     6121 gcttggtaca tgcagttctc tttgcgaaga tacaccactg tggccacaga actaaagtag
     6181 tccggcttgt cgccttggcc caggttctcg gatttgacct catacaaggt tttccagttg
     6241 gtgttactcc ctccgactcc gccgctcttt agatcagaga tggaaacacc atctaaggct
     6301 tgtccttctg cgtcaaacca tccacgaagc ttataggcct ctgggatgtc aggattcgca
     6361 atgatagtgc ttgaagacag cacggagagg ctccgtccac cgaaatcaga gactcgggct
     6421 cctttgatag ccaacacggg ctgtctagag gggaattgtt atccgctcac aattccccta
     6481 tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg gacgcatcgt
     6541 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     6601 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     6661 ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc
     6721 ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca
     6781 taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg
     6841 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     6901 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     6961 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     7021 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     7081 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     7141 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     7201 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     7261 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt c
//
