LOCUS       sequence-416910-        5526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of GFP-tagged hsRPA2-Ala10 in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5526)
  TITLE     Wold lab plasmids 2023
REFERENCE   2  (bases 1 to 5526)
  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: pEGFP-hsRPA32-Ala10 (MSW#508)
FEATURES             Location/Qualifiers
     source          1..5526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(576..1286,1287..1289,1290..1292))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(576..1286,1287..1289,1290..1292))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(618..639)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     967..986
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1226..1247
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1289..1298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1337..1540)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1366..1386)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1541..1844
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2019..2607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2099..2118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2936..2983
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3008..3027
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3215..4009)
                     /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             complement(3215..4009)
                     /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(3326..3345)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3936..3955
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4044..4401)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4058..4193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4210..4401
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4358..4378
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4403..4507)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4534..4989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4621..4640)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4831..4852
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4996..5117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5007..5026)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5061..5080
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ataggtgctc tccctgctga gggctggctg ttggctttgc ttagtaccat gtgtgcattg
       61 atcacttcca gaatatgtgt ggtgaactca ttcatatcct ccaggggcat gatcttaaag
      121 gctaccaggc tctttttgtt ctgaaaagat ctcaggtggc ctgccacttt cacatatgtt
      181 tctggaggaa ccacagtgtt ttcactgctg gtgtcatctg tgtcaaccca ctggcgaacg
      241 tccatgggtg cagctgtcat gtcatctatt ttgtaaacaa tgttggttgg agccttctct
      301 gcatgtctga tgatccccac aatagtgacc tgtgaaatct caacattccc aattctgaac
      361 acttcatcaa ccaaagtggc agaaagcagc tgagatatag tacagggcac aatgtgctgg
      421 gctcgggctc ttgctttctt ttcggcttga gcaggtgcgg gcgctccaaa gccccccggc
      481 gcctgcgcgt agccgccggc tcccccgtat gccgcggcgc cataggcttc gaatccagca
      541 ttccacatgc ctccacctcg agatctgagt ccggacttgt acagctcgtc catgccgaga
      601 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
      661 tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
      721 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
      781 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
      841 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
      901 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
      961 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     1021 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
     1081 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     1141 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     1201 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     1261 ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtagcgc tagcggatct
     1321 gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     1381 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     1441 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     1501 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     1561 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     1621 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     1681 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     1741 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     1801 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     1861 ggctatgaac taatgacccc gtaattgatt actattaata actaatgcat ggcggtaata
     1921 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1981 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2041 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2101 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2161 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2221 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2281 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2341 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2401 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2461 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2521 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2581 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2641 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2701 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2761 taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca
     2821 cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg gccccaatgg
     2881 ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca
     2941 aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc gcgggttcct
     3001 tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc gaagaactcc
     3061 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
     3121 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc
     3181 gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     3241 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     3301 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     3361 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     3421 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     3481 tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     3541 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     3601 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     3661 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     3721 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     3781 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca
     3841 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     3901 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     3961 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     4021 ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac
     4081 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt
     4141 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
     4201 gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     4261 gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc
     4321 atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac
     4381 agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc atttatcagg
     4441 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4501 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     4561 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4621 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4681 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4741 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4801 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4861 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4921 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4981 ttacaattta cgcgttaaga tacattgatg agtttggaca aaccacaact agaatgcagt
     5041 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     5101 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     5161 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     5221 atcagttatc tagatccggt ggatcccggg cccgcggtac cttattctgc atctgtggat
     5281 ttaaaatggt catcatccac agtagaatag atgtgcccct cattgctcag aaaatccaca
     5341 gcttgcttga ttgaggatac agacatgtgt ttcagctggt tcttgagatc ctgaaagttc
     5401 aacccttcag gtcttggaca agccttaatc aaattcaaca cctggttttg ggccacagtg
     5461 aggccatttg ctggcatgaa gctattccca ccaaagttcc ctgcttcact cattcctgga
     5521 ttgctg
//
