LOCUS       sequence-420345-        8898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human RPA70 N-terminally fused to mVenus and
            RPA32 and RPA14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8898)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 8898)
  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: pmVen-RPA70-IRES-RPA32-IRES-RPA14
FEATURES             Location/Qualifiers
     source          1..8898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    159..206
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      535..1123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1024..1043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1298..1601
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1602..1805
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1756..1776
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1844..1853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1850..1852,1853..1855,1856..2566)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1895..1916)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2156..2175)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2503..2524
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6923..7044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6960..6979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7014..7033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(7051..7506)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7188..7209)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7400..7419
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7533..7637
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7639..7996
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7639..7830
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7662..7682)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7847..7982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8031..8825
                     /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             8031..8825
                     /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(8085..8104)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8695..8714
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
       61 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
      121 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
      181 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
      241 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
      301 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
      361 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
      421 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
      481 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
      541 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
      601 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
      661 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
      721 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
      781 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
      841 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
      901 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
      961 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1021 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1081 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     1141 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     1201 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     1261 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1321 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1381 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     1441 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     1501 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     1561 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     1621 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     1681 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     1741 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     1801 gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa
     1861 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1921 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1981 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2041 cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     2101 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2161 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2221 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2281 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     2341 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     2401 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     2461 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2521 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc
     2581 tcgagctcgt atggtcggcc agctgagcga gggggccatt gcggccatca tgcagaaggg
     2641 ggatacaaac ataaagccca tcctccaagt catcaacatc cgtcccatta ctacggggaa
     2701 tagtccgccg cgttatcgac tgctcatgag tgatggattg aacactctat cctctttcat
     2761 gttggcgaca cagttgaacc ctctcgtgga ggaagaacaa ttgtccagca actgtgtatg
     2821 ccagattcac agatttattg tgaacactct gaaagacgga aggagagtag ttatcttgat
     2881 ggaattagaa gttttgaagt cagctgaagc agttggagtg aagattggca atccagtgcc
     2941 ctataatgaa ggactcgggc agccgcaagt agctcctcca gcgccagcag ccagcccagc
     3001 agcaagcagc aggccccagc cgcagaatgg aagctcggga atgggttcta ctgtttctaa
     3061 ggcttatggt gcttcaaaga catttggaaa agctgcaggt cccagcctgt cacacacttc
     3121 tgggggaaca cagtccaaag tggtgcccat tgccagcctc actccttacc agtccaagtg
     3181 gaccatttgt gctcgtgtta ccaacaaaag tcagatccgt acctggagca actcccgagg
     3241 ggaagggaag cttttctccc tagaactggt tgacgaaagt ggtgaaatcc gagctacagc
     3301 tttcaatgag caagtggaca agttctttcc tcttattgaa gtgaacaagg tgtattattt
     3361 ctcgaaaggc accctgaaga ttgctaacaa gcagttcaca gctgttaaaa atgactacga
     3421 gatgaccttc aataacgaga cttccgtcat gccctgtgag gacgaccatc atttacctac
     3481 ggttcagttt gatttcacgg ggattgatga cctcgagaac aagtcgaaag actcacttgt
     3541 agacatcatc gggatctgca agagctatga agacgccact aaaatcacag tgaggtctaa
     3601 caacagagaa gttgccaaga ggaatatcta cttgatggac acatctggga aggtggtgac
     3661 tgctacactg tggggggaag atgctgataa atttgatggt tctagacagc ccgtgttggc
     3721 tatcaaagga gcccgagtct ctgatttcgg tggacggagc ctctccgtgc tgtcttcaag
     3781 cactatcatt gcgaatcctg acatcccaga ggcctataag cttcgtggat ggtttgacgc
     3841 agaaggacaa gccttagatg gtgtttccat ctctgatcta aagagcggcg gagtcggagg
     3901 gagtaacacc aactggaaaa ccttgtatga ggtcaaatcc gagaacctgg gccaaggcga
     3961 caagccggac tactttagtt ctgtggccac agtggtgtat cttcgcaaag agaactgcat
     4021 gtaccaagcc tgcccgactc aggactgcaa taagaaagtg attgatcaac agaatggatt
     4081 gtaccgctgt gagaagtgcg acaccgaatt tcccaatttc aagtaccgca tgatcctgtc
     4141 agtaaatatt gcagattttc aagagaatca gtgggtgact tgtttccagg agtctgctga
     4201 agctatcctt ggacaaaatg ctgcttatct tggggaatta aaagacaaga atgaacaggc
     4261 atttgaagaa gttttccaga atgccaactt ccgatctttc atattcagag tcagggtcaa
     4321 agtggagacc tacaacgacg agtctcgaat taaggccact gtgatggacg tgaagcccgt
     4381 ggactacaga gagtatggcc gaaggctggt catgagcatc aggagaagtg cattgatgtg
     4441 aggatctttc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     4501 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     4561 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     4621 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     4681 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     4741 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     4801 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     4861 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     4921 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     4981 ccgaaccacg gggacgtggt tttcctttga aaaacacgat aagaattctg ccaccatgtg
     5041 gaacagtgga ttcgaaagct atggcagctc ctcatacggg ggagccggcg gctacacgca
     5101 gtccccgggg ggctttggat cgcccgcacc ttctcaagcc gaaaagaaat caagagcccg
     5161 agcccagcac attgtgccct gtactatatc tcagctgctt tctgccactt tggttgatga
     5221 agtgttcaga attgggaatg ttgagatttc acaggtcact attgtgggga tcatcagaca
     5281 tgcagagaag gctccaacca acattgttta caaaatagat gacatgacag ctgcacccat
     5341 ggacgttcgc cagtgggttg acacagatga caccagcagt gaaaacactg tggttcctcc
     5401 agaaacatat gtgaaagtgg caggccacct gagatctttt cagaacaaaa agagcctggt
     5461 agcctttaag atcatgcccc tggaggatat gaatgagttc accacacata ttctggaagt
     5521 gatcaatgca cacatggtac taagcaaagc caacagccag ccctcagcag ggagagcacc
     5581 tatcagcaat ccaggaatga gtgaagcagg gaactttggt gggaatagct tcatgccagc
     5641 aaatggcctc actgtggccc aaaaccaggt gttgaatttg attaaggctt gtccaagacc
     5701 tgaagggttg aactttcagg atctcaagaa ccagctgaaa cacatgtctg tatcctcaat
     5761 caagcaagct gtggattttc tgagcaatga ggggcacatc tattctactg tggatgatga
     5821 ccattttaaa tccacagatg cagaataagg atccaattcc gcccctctcc ctcccccccc
     5881 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     5941 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     6001 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     6061 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
     6121 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     6181 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     6241 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     6301 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
     6361 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     6421 aaaacacgat aaagatctca tggtggacat gatggacttg cccaggtcgc gcatcaacgc
     6481 cggcatgcta gctcaattca tcgacaagcc tgtctgcttc gtagggaggc tggaaaagat
     6541 tcatcccacc ggaaaaatgt ttattctttc agatggagaa ggaaaaaatg gaaccatcga
     6601 gttgatggaa ccccttgatg aagaaatctc tggaattgtg gaagtggttg gaagagtaac
     6661 cgccaaggcc accatcttgt gtacatctta tgtccagttt aaagaagata gccatccttt
     6721 tgatcttgga ctttacaatg aagctgtgaa aattatccat gacttccctc agttttatcc
     6781 tttagggatt gtgcaacatg attgatctag ataactgatc ataatcagcc ataccacatt
     6841 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     6901 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     6961 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7021 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     7081 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     7141 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     7201 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     7261 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     7321 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     7381 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     7441 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     7501 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7561 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7621 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     7681 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     7741 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     7801 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     7861 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     7921 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     7981 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     8041 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     8101 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     8161 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     8221 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     8281 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     8341 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     8401 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     8461 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     8521 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     8581 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     8641 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     8701 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     8761 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     8821 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     8881 agatttcgat tccaccgc
//
