LOCUS       sequence-420528-        8168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human RPA70 and RPA32 and RPA14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8168)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 8168)
  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"
COMMENT     Sequence Label: pRPA70-IRES-RPA32-IRES-RPA14
FEATURES             Location/Qualifiers
     source          1..8168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    4960..5081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4997..5016)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5051..5070
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5088..5543)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5225..5246)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5437..5456
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5570..5674
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5676..6033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5676..5867
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5699..5719)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5884..6019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6068..6862
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6068..6862
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6122..6141)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6732..6751
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7094..7141
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7470..8058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7959..7978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gctaccggac tcagatctcg agctcgtatg gtcggccagc tgagcgaggg ggccattgcg
      661 gccatcatgc agaaggggga tacaaacata aagcccatcc tccaagtcat caacatccgt
      721 cccattacta cggggaatag tccgccgcgt tatcgactgc tcatgagtga tggattgaac
      781 actctatcct ctttcatgtt ggcgacacag ttgaaccctc tcgtggagga agaacaattg
      841 tccagcaact gtgtatgcca gattcacaga tttattgtga acactctgaa agacggaagg
      901 agagtagtta tcttgatgga attagaagtt ttgaagtcag ctgaagcagt tggagtgaag
      961 attggcaatc cagtgcccta taatgaagga ctcgggcagc cgcaagtagc tcctccagcg
     1021 ccagcagcca gcccagcagc aagcagcagg ccccagccgc agaatggaag ctcgggaatg
     1081 ggttctactg tttctaaggc ttatggtgct tcaaagacat ttggaaaagc tgcaggtccc
     1141 agcctgtcac acacttctgg gggaacacag tccaaagtgg tgcccattgc cagcctcact
     1201 ccttaccagt ccaagtggac catttgtgct cgtgttacca acaaaagtca gatccgtacc
     1261 tggagcaact cccgagggga agggaagctt ttctccctag aactggttga cgaaagtggt
     1321 gaaatccgag ctacagcttt caatgagcaa gtggacaagt tctttcctct tattgaagtg
     1381 aacaaggtgt attatttctc gaaaggcacc ctgaagattg ctaacaagca gttcacagct
     1441 gttaaaaatg actacgagat gaccttcaat aacgagactt ccgtcatgcc ctgtgaggac
     1501 gaccatcatt tacctacggt tcagtttgat ttcacgggga ttgatgacct cgagaacaag
     1561 tcgaaagact cacttgtaga catcatcggg atctgcaaga gctatgaaga cgccactaaa
     1621 atcacagtga ggtctaacaa cagagaagtt gccaagagga atatctactt gatggacaca
     1681 tctgggaagg tggtgactgc tacactgtgg ggggaagatg ctgataaatt tgatggttct
     1741 agacagcccg tgttggctat caaaggagcc cgagtctctg atttcggtgg acggagcctc
     1801 tccgtgctgt cttcaagcac tatcattgcg aatcctgaca tcccagaggc ctataagctt
     1861 cgtggatggt ttgacgcaga aggacaagcc ttagatggtg tttccatctc tgatctaaag
     1921 agcggcggag tcggagggag taacaccaac tggaaaacct tgtatgaggt caaatccgag
     1981 aacctgggcc aaggcgacaa gccggactac tttagttctg tggccacagt ggtgtatctt
     2041 cgcaaagaga actgcatgta ccaagcctgc ccgactcagg actgcaataa gaaagtgatt
     2101 gatcaacaga atggattgta ccgctgtgag aagtgcgaca ccgaatttcc caatttcaag
     2161 taccgcatga tcctgtcagt aaatattgca gattttcaag agaatcagtg ggtgacttgt
     2221 ttccaggagt ctgctgaagc tatccttgga caaaatgctg cttatcttgg ggaattaaaa
     2281 gacaagaatg aacaggcatt tgaagaagtt ttccagaatg ccaacttccg atctttcata
     2341 ttcagagtca gggtcaaagt ggagacctac aacgacgagt ctcgaattaa ggccactgtg
     2401 atggacgtga agcccgtgga ctacagagag tatggccgaa ggctggtcat gagcatcagg
     2461 agaagtgcat tgatgtgagg atctttcgcc cctctccctc ccccccccct aacgttactg
     2521 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     2581 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     2641 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     2701 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     2761 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     2821 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     2881 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     2941 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     3001 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataag
     3061 aattctgcca ccatgtggaa cagtggattc gaaagctatg gcagctcctc atacggggga
     3121 gccggcggct acacgcagtc cccggggggc tttggatcgc ccgcaccttc tcaagccgaa
     3181 aagaaatcaa gagcccgagc ccagcacatt gtgccctgta ctatatctca gctgctttct
     3241 gccactttgg ttgatgaagt gttcagaatt gggaatgttg agatttcaca ggtcactatt
     3301 gtggggatca tcagacatgc agagaaggct ccaaccaaca ttgtttacaa aatagatgac
     3361 atgacagctg cacccatgga cgttcgccag tgggttgaca cagatgacac cagcagtgaa
     3421 aacactgtgg ttcctccaga aacatatgtg aaagtggcag gccacctgag atcttttcag
     3481 aacaaaaaga gcctggtagc ctttaagatc atgcccctgg aggatatgaa tgagttcacc
     3541 acacatattc tggaagtgat caatgcacac atggtactaa gcaaagccaa cagccagccc
     3601 tcagcaggga gagcacctat cagcaatcca ggaatgagtg aagcagggaa ctttggtggg
     3661 aatagcttca tgccagcaaa tggcctcact gtggcccaaa accaggtgtt gaatttgatt
     3721 aaggcttgtc caagacctga agggttgaac tttcaggatc tcaagaacca gctgaaacac
     3781 atgtctgtat cctcaatcaa gcaagctgtg gattttctga gcaatgaggg gcacatctat
     3841 tctactgtgg atgatgacca ttttaaatcc acagatgcag aataaggatc caattccgcc
     3901 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     3961 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     4021 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     4081 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     4141 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     4201 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     4261 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     4321 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     4381 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     4441 acgtggtttt cctttgaaaa acacgataaa gatctcatgg tggacatgat ggacttgccc
     4501 aggtcgcgca tcaacgccgg catgctagct caattcatcg acaagcctgt ctgcttcgta
     4561 gggaggctgg aaaagattca tcccaccgga aaaatgttta ttctttcaga tggagaagga
     4621 aaaaatggaa ccatcgagtt gatggaaccc cttgatgaag aaatctctgg aattgtggaa
     4681 gtggttggaa gagtaaccgc caaggccacc atcttgtgta catcttatgt ccagtttaaa
     4741 gaagatagcc atccttttga tcttggactt tacaatgaag ctgtgaaaat tatccatgac
     4801 ttccctcagt tttatccttt agggattgtg caacatgatt gatctagata actgatcata
     4861 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4921 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4981 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5041 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     5101 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5161 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     5221 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5281 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5341 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     5401 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     5461 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5521 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5581 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5641 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     5701 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5761 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5821 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5881 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5941 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6001 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     6061 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     6121 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     6181 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     6241 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     6301 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     6361 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     6421 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     6481 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6541 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     6601 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6661 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6721 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6781 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6841 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6901 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6961 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     7021 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     7081 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     7141 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     7201 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     7261 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     7321 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7381 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7441 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7501 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7561 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7621 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7681 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7741 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7801 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7861 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7921 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7981 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8041 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8101 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8161 accgccat
//
