LOCUS       sequence-468353-        6701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged wild type human RAD52 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6701)
  AUTHORS   Jalan M, Sharma A, Pei X, Weinhold N, Buechelmaier ES, Zhu Y, 
            Ahmed-Seghir S, Ratnakumar A, Di Bona M, McDermott N, Gomez-Aguilar 
            J, Anderson KS, Ng CKY, Selenica P, Bakhoum SF, Reis-Filho JS, Riaz 
            N, Powell SN
  TITLE     RAD52 resolves transcription-replication conflicts to mitigate 
            R-loop induced genome instability.
  JOURNAL   Nat Commun. 2024 Sep 5;15(1):7776. doi: 10.1038/s41467-024-51784-x.
  PUBMED    39237529
REFERENCE   2  (bases 1 to 6701)
  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"; journalName: "Nat 
            Commun."; date: "2024-09-5"; volume: "15(1):7776. doi"; pages: " 
            10.1038/s41467-024-51784-x"
COMMENT     Sequence Label: pcDNA3.1-HA-RAD52-WT
FEATURES             Location/Qualifiers
     source          1..6701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             926..952
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2292..2309)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2298..2522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2568..2996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2655..2674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2865..2886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3005..3025)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3010..3339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3190..3325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3252..3271
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3406..4200
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3460..3479)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4070..4089
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4374..4495
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4411..4430)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4465..4484
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4544..4560)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4544..4560)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4557..4579)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4568..4584
                     /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)."
     promoter        complement(join(4592..4598,4599..4616,4617..4622))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4637..4658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4775..4792)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4946..5534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5026..5045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5705..6496,6497..6565))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6328..6347
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6566..6670)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgtaccc atacgatgtt ccagattacg ctggtaccat
      961 gtctgggact gaggaagcaa ttcttggagg acgtgacagc catcctgctg ctggcggcgg
     1021 ctcagtgtta tgctttggac agtgccagta cacagcagaa gagtaccagg ccatccagaa
     1081 ggccctgagg cagaggctgg gcccagaata cataagtagc cgcatggctg gcggaggcca
     1141 gaaggtgtgc tacattgagg gtcatcgggt aattaatctg gccaatgaga tgtttggtta
     1201 caatggctgg gcacactcca tcacgcagca gaatgtggat tttgttgacc tcaacaatgg
     1261 caagttctac gtgggagtct gtgcatttgt gagggtccag ctgaaggatg gttcatatca
     1321 tgaagatgtt ggttatggtg ttagtgaggg cctcaagtcc aaggctttat ctttggagaa
     1381 ggcaaggaag gaggcggtga cagacgggct gaagcgagcc ctcaggagtt ttgggaatgc
     1441 acttggaaac tgtattctgg acaaagacta cctgagatca ctaaataagc ttccacgcca
     1501 gttgcctctt gaagtggatt taactaaagc gaagagacaa gatcttgaac cgtctgtgga
     1561 ggaggcaaga tacaacagct gccgaccgaa catggccctg ggacacccac agctgcagca
     1621 ggtgacctcc ccttccagac ccagccatgc tgtgataccg gcggaccagg actgcagctc
     1681 ccgaagcctg agctcatccg ccgtggagag cgaggccacg caccagcgga agctccggca
     1741 gaagcagctg cagcagcagt tccgggagcg gatggagaag cagcaggttc gagtctccac
     1801 gccgtcagct gagaagagtg aggcagcgcc tccggcccct cctgtgacgc acagcactcc
     1861 tgtaactgtc tcagaaccac tcctggagaa agacttcctt gcaggagtga ctcaagaatt
     1921 aatcaagact cttgaagaca actctgaaaa gtgggctgtg actcccgatg caggggatgg
     1981 tgtggtcaag ccctcgtcta gagcagaccc agcccagacc tctgacacat tagccttgaa
     2041 caaccagatg gtgacccaga acaggactcc acacagcgtt tgccaccaga aaccacaagc
     2101 aaaatctgga tcttgggacc tccaaactta tagcgctgac caacgcacaa caggaaactg
     2161 ggaatctcat aggaagagcc aggacatgaa gaaaaggaaa tatgatccat cttaaggatc
     2221 cgaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac
     2281 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2341 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2401 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2461 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2521 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2581 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2641 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2701 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2761 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2821 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2881 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2941 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3001 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3061 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3241 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3361 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3421 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3481 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3541 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3601 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3661 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3721 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3781 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3841 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3901 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3961 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4021 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4081 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4141 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4201 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4261 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4321 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4381 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4441 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4501 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4561 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4621 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4681 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4741 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4801 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4861 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4921 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4981 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5041 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5101 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5161 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5221 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5281 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5341 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5401 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5461 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5521 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5581 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5641 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5701 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5761 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5821 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5881 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5941 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6001 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6061 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6121 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6181 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6241 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6301 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6361 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6421 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6481 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6541 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6601 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6661 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
