LOCUS       sequence-468354-        6374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged mutant human RAD52 in mammalian cells with a 
            C-terminal deletion (Delta-302-410 amino acids)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6374)
  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 6374)
  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-∆C
FEATURES             Location/Qualifiers
     source          1..6374
                     /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(1965..1982)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1971..2195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2241..2669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2328..2347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2538..2559
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2678..2698)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2683..3012
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2863..2998
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2925..2944
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3079..3873
                     /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(3133..3152)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3743..3762
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4047..4168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4084..4103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4138..4157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4217..4233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4217..4233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4230..4252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4241..4257
                     /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(4265..4271,4272..4289,4290..4295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4310..4331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4448..4465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4619..5207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4699..4718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5378..6169,6170..6238))
                     /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     6001..6020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6239..6343)
                     /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 acagcgacat
     1861 gaagaaaagg aaatatgatc catcttaagg atccgaattc tgcagatatc cagcacagtg
     1921 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     1981 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2041 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2101 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2161 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2221 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2461 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2521 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2581 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2641 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2701 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2761 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2821 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2881 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2941 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3001 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3061 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3121 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3181 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3241 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3301 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3361 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3421 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3481 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3541 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3601 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3661 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3721 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3781 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3841 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3901 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3961 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4021 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4081 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4141 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4201 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4261 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4321 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4381 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4441 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4501 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4561 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4621 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4681 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4741 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4801 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4861 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4921 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4981 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5041 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5101 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5161 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5221 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5281 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5341 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5401 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5461 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5521 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5581 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5641 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5701 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5761 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5821 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5881 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5941 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6001 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6061 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6121 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6181 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6241 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6301 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6361 tgccacctga cgtc
//
