LOCUS       sequence-420413-        6471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human RPA32 N-terminally fused to mVenus and
            RPA14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6471)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 6471)
  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: pVen-RPA32-IRES-RPA14
FEATURES             Location/Qualifiers
     source          1..6471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        370..573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     524..544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      612..621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(618..620,621..623,624..1334)
                     /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(663..684)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(924..943)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1271..1292
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2370..2387)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2597..2616
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     polyA_signal    3264..3385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3301..3320)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3355..3374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3392..3847)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3529..3550)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3741..3760
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3874..3978
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3980..4337
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3980..4171
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4003..4023)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4188..4323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4372..5166
                     /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             4372..5166
                     /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(4426..4445)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5036..5055
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5354..5373)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5398..5445
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5774..6362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6263..6282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      601 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      661 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      721 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
      781 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct
      841 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      901 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      961 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1021 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     1081 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     1141 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1201 tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga
     1261 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1321 acgagctgta caagtccgga ctcagatctc gagctcaagc ttcgaattct gccaccatgt
     1381 ggaacagtgg attcgaaagc tatggcagct cctcatacgg gggagccggc ggctacacgc
     1441 agtccccggg gggctttgga tcgcccgcac cttctcaagc cgaaaagaaa tcaagagccc
     1501 gagcccagca cattgtgccc tgtactatat ctcagctgct ttctgccact ttggttgatg
     1561 aagtgttcag aattgggaat gttgagattt cacaggtcac tattgtgggg atcatcagac
     1621 atgcagagaa ggctccaacc aacattgttt acaaaataga tgacatgaca gctgcaccca
     1681 tggacgttcg ccagtgggtt gacacagatg acaccagcag tgaaaacact gtggttcctc
     1741 cagaaacata tgtgaaagtg gcaggccacc tgagatcttt tcagaacaaa aagagcctgg
     1801 tagcctttaa gatcatgccc ctggaggata tgaatgagtt caccacacat attctggaag
     1861 tgatcaatgc acacatggta ctaagcaaag ccaacagcca gccctcagca gggagagcac
     1921 ctatcagcaa tccaggaatg agtgaagcag ggaactttgg tgggaatagc ttcatgccag
     1981 caaatggcct cactgtggcc caaaaccagg tgttgaattt gattaaggct tgtccaagac
     2041 ctgaagggtt gaactttcag gatctcaaga accagctgaa acacatgtct gtatcctcaa
     2101 tcaagcaagc tgtggatttt ctgagcaatg aggggcacat ctattctact gtggatgatg
     2161 accattttaa atccacagat gcagaataag gatccaattc cgcccctctc cctccccccc
     2221 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     2281 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     2341 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     2401 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     2461 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     2521 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     2581 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     2641 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     2701 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     2761 aaaaacacga taaagatctc atggtggaca tgatggactt gcccaggtcg cgcatcaacg
     2821 ccggcatgct agctcaattc atcgacaagc ctgtctgctt cgtagggagg ctggaaaaga
     2881 ttcatcccac cggaaaaatg tttattcttt cagatggaga aggaaaaaat ggaaccatcg
     2941 agttgatgga accccttgat gaagaaatct ctggaattgt ggaagtggtt ggaagagtaa
     3001 ccgccaaggc caccatcttg tgtacatctt atgtccagtt taaagaagat agccatcctt
     3061 ttgatcttgg actttacaat gaagctgtga aaattatcca tgacttccct cagttttatc
     3121 ctttagggat tgtgcaacat gattgatcta gataactgat cataatcagc cataccacat
     3181 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3241 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3301 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3361 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     3421 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3481 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3541 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3601 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3661 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3721 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3781 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3841 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3901 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3961 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4021 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4081 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4141 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4201 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4261 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4321 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4381 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4441 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4501 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4561 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4621 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4681 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4741 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4801 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4861 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4921 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4981 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5041 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5101 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5161 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5221 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5281 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5341 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5401 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5461 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5521 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5581 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5641 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5701 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5761 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5821 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5881 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5941 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6001 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6061 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6121 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6181 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6241 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6301 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6361 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6421 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc a
//
