LOCUS       sequence-413701-        8882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Alternative human Replication Protein A (RPA1,4,3) in 
            E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8882)
  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 8882)
  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: p11d-aRPA (MSW#712)
FEATURES             Location/Qualifiers
     source          1..8882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2068..2259
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2253..2275)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2361..2500
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2416..2435
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2515..2532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2686..3274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2766..2785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3445..4236,4237..4305))
                     /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     4068..4087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4306..4410)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4441..4487)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     RBS             5284..5306
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             5702..5724
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             8090..8112
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    8127..8151
                     /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(8151..8170)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8152..8170)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8254..8273)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        8479..8556
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(8576..8595)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
       61 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
      121 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
      181 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
      241 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
      301 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
      361 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
      421 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
      481 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
      541 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
      601 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
      661 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      721 ggcacgacag tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
      781 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
      841 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
      901 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
      961 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     1021 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     1081 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg
     1141 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
     1201 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc
     1261 ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
     1321 ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg
     1381 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
     1441 cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga
     1501 actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc
     1561 agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg
     1621 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa
     1681 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca
     1741 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg
     1801 ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca
     1861 cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc
     1921 cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca
     1981 tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac
     2041 agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg
     2101 gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg
     2161 gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc
     2221 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     2281 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     2341 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     2401 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat tgcggtgtga
     2461 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     2521 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2581 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2641 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2701 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2761 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2821 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2881 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2941 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3001 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3061 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3121 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3181 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3241 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3301 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3361 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3421 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3481 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3541 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3601 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3661 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3721 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     3781 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3841 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3901 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3961 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4021 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     4081 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4141 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4201 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4261 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4321 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4381 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4441 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatta tcaatcagca
     4501 gacttaaaat gctcccgatc cacagtggga tagatgtggc cctcaacggt cagataatca
     4561 atcgcttcct tgatggcctt gacgctaagg tcgcagagct gagcccggag ctcatggatg
     4621 ctcttccctt cctgatgagg acactcatga atcaaacgca gcacttcgtc ctggatgaaa
     4681 ttgcggtgac tctcatcgtt atccccagca tcattcactt ctgatggaga cacaggcaca
     4741 ctttctacag tggtatcacg acgggcttta tccagcatca tgtgtgcatt gaccgtttcc
     4801 agaatatgca cggtgaactc gttcatgtcc tctaggacat gaattttcaa tacctcaagg
     4861 ctctttgttc ccgtgggaca tttgaggata ccaaacactt tgacatatac tccgactgac
     4921 aatggagtca cctgcttgac tttctctcta ccaaaccact gtcgggcctc gattggtttc
     4981 gcggtcatat catcaatttt gtaacaaatg tgatttgaag ccttctctgc ccctctgatt
     5041 acccccacga tggagacctg ggaaactata attcccctaa ccttgaacac agggtcaaac
     5101 acagtagagc tgagtagctg gttcacgtta cacggtacaa cgtcctgaat tcggacctta
     5161 ggtctttggg tcttaatagc aggagttgca tctctctcac acagttggtc actgcctcca
     5221 ctcgctccat cagcagcaga aatgctgcca tagctcccaa acccactctt actcatatgt
     5281 atatctcctt cttaaagtta aacaaaatta tttctaggat ccatcaagat caatcatgtt
     5341 gcacaatccc taaaggataa aactgaggga agtcatggat aattttcaca gcttcattgt
     5401 aaagtccaag atcaaaagga tggctatctt ctttaaactg gacataagat gtacacaaga
     5461 tggtggcctt ggcggttact cttccaacca cttccacaat tccagagatt tcttcatcaa
     5521 ggggttccat caactcgatg gttccatttt ttccttctcc atctgaaaga ataaacattt
     5581 ttccggtggg atgaatcttt tccagcctcc ctacgaagca gacaggcttg tcgatgaatt
     5641 gagctagcat gccggcgttg atgcgcgacc tgggcaagtc catcatgtcc accatggtat
     5701 atctccttct taaagttaaa caaaattatt tctaggatcc cccgggctgc aggaattcta
     5761 ggaagacaca cttactaaaa tccccaggtg ctcatgaccg cttctcagct cgccccgctg
     5821 agaagaaccc aagttgtgag cggggatggg ggggggtggt caccactgga gaagcaaaaa
     5881 cctagttaca taatttactt catggtctgc agttagggtc agtgacttac gacataattc
     5941 ctgcttgatg ataatgaaat tgacagaagc ctgaaggcag agggagtgac tggtcttgcg
     6001 ctagccactg gctgactccg taggcctgac acagtctaca ccacaaccac ctgagcgtat
     6061 tttcctttgc ctaccgatgg gttctgagat gcgcacgagg gaggaaattg cttagtccac
     6121 catcaagagc ctctgcactg tgtagctaac atgggatcgt cacacggagg tgggaggaaa
     6181 tcgattccat tctgcctatt tgcaaacttc tgcccgattg gcactgctcc tctcacatca
     6241 atgcacttct cctgatgctc atgaccagcc ttcggccata ctctctgtag tccacgggct
     6301 tcacgtccat cacagtggcc ttaattcgag actcgtcgtt gtaggtctcc actttgaccc
     6361 tgactctgaa tatgaaagat cggaagttgg cattctggaa aacttcttca aatgcctgtt
     6421 cattcttgtc ttttaattcc ccaagataag cagcattttg tccaaggata gcttcagcag
     6481 actcctggaa acaagtcacc cactgattct cttgaaaatc tgcaatattt actgacagga
     6541 tcatgcggta cttgaaattg ggaaattcgg tgtcgcactt ctcacagcgg tacaatccat
     6601 tctgttgatc aatcactttc ttattgcagt cctgagtcgg gcaggcttgg tacatgcagt
     6661 tctctttgcg aagatacacc actgtggcca cagaactaaa gtagtccggc ttgtcgcctt
     6721 ggcccaggtt ctcggatttg acctcataca aggttttcca gttggtgtta ctccctccga
     6781 ctccgccgct ctttagatca gagatggaaa caccatctaa ggcttgtcct tctgcgtcaa
     6841 accatccacg aagcttatag gcctctggga tgtcaggatt cgcaatgata gtgcttgaag
     6901 acagcacgga gaggctccgt ccaccgaaat cagagactcg ggctcctttg atagccaaca
     6961 cgggctgtct agaaccatca aatttatcag catcttcccc ccacagtgta gcagtcacca
     7021 ccttcccgga tgtgtccatc aagtagatat tcctcttggc aacttctctg ttgttagacc
     7081 tcactgtgat tttagtggcg tcttcatagc tcttgcagat cccgatgatg tctacaagtg
     7141 agtctttcga cttgttctcg aggtcatcaa tccccgtgaa atcaaactga accgtaggta
     7201 aatgatggtc gtcctcacag ggcatgacgg aagtctcgtt attgaaggtc atctcgtagt
     7261 catttttaac agctgtgaac tgcttgttag caatcttcag ggtgcctttc gagaaataat
     7321 acaccttgtt cacttcaata agaggaaaga acttgtccac ttgctcattg aaagctgtag
     7381 ctcggatttc accactttcg tcaaccagtt ctagggagaa aagcttccct tcccctcggg
     7441 agttgctcca ggtacggatc tgacttttgt tggtaacacg agcacaaatg gtccacttgg
     7501 actggtaagg agtgaggctg gcaatgggca ccactttgga ctgtgttccc ccagaagtgt
     7561 gtgacaggct gggacctgca gcttttccaa atgtctttga agcaccataa gccttagaaa
     7621 cagtagaacc cattcccgag cttccattct gcggctgggg cctgctgctt gctgctgggc
     7681 tggctgctgg cgctggagga gctacttgcg gctgcccgag tccttcatta tagggcactg
     7741 gattgccaat cttcactcca actgcttcag ctgacttcaa aacttctaat tccatcaaga
     7801 taactactct ccttccgtct ttcagagtgt tcacaataaa tctgtgaatc tggcatacac
     7861 agttgctgga caattgttct tcctccacga gagggttcaa ctgtgtcgcc aacatgaaag
     7921 aggatagagt gttcaatcca tcactcatga gcagtcgata acgcggcgga ctattccccg
     7981 tagtaatggg acggatgttg atgacttgga ggatgggctt tatgtttgta tcccccttct
     8041 gcatgatggc cgcaatggcc ccctcgctca gctggccgac catggtatat ctccttctta
     8101 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg
     8161 agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc atcgtggccg
     8221 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     8281 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     8341 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg
     8401 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
     8461 gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
     8521 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
     8581 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     8641 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     8701 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     8761 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     8821 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     8881 gc
//
