LOCUS       sequence-395051-        6971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a mutant form of human RAD23B that lacks the XPC-binding 
            domain. Variant of plasmid pCMV6-AN-DDK_WT RAD23B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  AUTHORS   Ashton NW, Jaiswal N, Cestari Moreno N, Semenova IV, D'Orlando DA, 
            Teatin Latancia M, McIntyre J, Woodgate R, Bezsonova I
  TITLE     A novel interaction between RAD23A/B and Y-family DNA polymerases.
  JOURNAL   J Mol Biol. 2023 Nov 5:168353. doi: 10.1016/j.jmb.2023.168353.
  PUBMED    37935254
REFERENCE   2  (bases 1 to 6971)
  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: "J Mol Biol.
            2023 Nov 5:168353. doi: 10.1016/j.jmb.2023.168353."
COMMENT     Sequence Label: pCMV6-AN-DDK_WT RAD23B_delta aa 276-339 (delta...
FEATURES             Location/Qualifiers
     source          1..6971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1014..1023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1023..1046
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2176..2192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2203..2825
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2402..2421)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2849..2869)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2854..3183
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3034..3169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3250..4044
                     /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(3304..3323)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3914..3933
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4218..4339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4255..4274)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4309..4328
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4388..4404)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4401..4423)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4412..4428
                     /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(4436..4442,4443..4460,4461..4466))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4481..4502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4619..4636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4790..5378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4870..4889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5549..6340,6341..6409))
                     /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     6172..6191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6410..6514)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6541..6969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6628..6647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6838..6859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccacca
     1021 tggattacaa ggatgacgac gataaggcga tcgccatgga gcatatgcag gtgacactga
     1081 agacactgca gcagcagact tttaagattg acattgaccc cgaggagacc gtgaaggcac
     1141 tgaaggagaa aatcgagagc gaaaagggca aagacgcatt ccctgtggcc gggcagaagc
     1201 tgatctacgc cggaaaaatt ctgaacgacg ataccgctct gaaggaatac aagatcgatg
     1261 agaagaattt cgtggtcgtg atggtcacta agcctaaagc cgtgtccacc ccagctcccg
     1321 caaccacaca gcagtctgcc ccagctagta ctaccgccgt gacaagctcc acaactacca
     1381 cagtcgcaca ggcacctact ccagtgccag ctctggcacc cacatcaact cctgccagca
     1441 tcacaccagc cagcgctact gcatctagtg aacctgcccc agcttccgcc gctaagcagg
     1501 agaaaccagc tgaaaaaccc gcagagactc ctgtggccac ctcacccacc gctacagaca
     1561 gcacctccgg cgattcaagc cggtccaacc tgttcgagga cgctacctcc gcactggtca
     1621 caggccagtc ttacgaaaat atggtgacag agatcatgtc tatggggtat gagcgggaac
     1681 aggtcattgc agccctgaga gctagtttta acaatcccga tagggcagtg gaatacctgc
     1741 tgatgggcat ccctggggac agagagtctc aggcagtcgt ggacccccct caggctgcaa
     1801 gtaccggagt cccacagtcc tctgcagtgg cagctgcagc agctactacc acagctacta
     1861 ccacaactac cagttcaggc gggcaccctg gaggaggagg aggcgggagt ggaggcatcg
     1921 cagaggccgg gtcaggacac atgaactaca tccaggtcac accacaggag aaggaagcaa
     1981 ttgaaaggct gaaagccctg ggattccccg agggcctggt catccaggcc tattttgctt
     2041 gcgagaagaa cgaaaatctg gcagccaact tcctgctgca gcagaatttt gacgaggatt
     2101 aagaattcgg atccacgcgt aagcttctcg agacgcgtta agcggccgca ctcgaggttt
     2161 aaacggccgg ccgcggtcat agctgtttcc tgaacagatc ccgggtggca tccctgtgac
     2221 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2281 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2341 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2401 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     2461 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     2521 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     2581 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     2641 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac
     2701 cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat
     2761 ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt
     2821 gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3001 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3061 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3121 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3181 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3241 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3301 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3361 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3421 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3481 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3541 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3601 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3661 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3721 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3781 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3841 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggccgaccgc
     3901 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3961 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4021 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4081 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4141 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4201 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4261 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4321 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4381 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4441 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4501 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4561 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4621 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4681 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4741 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4801 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4861 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4921 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4981 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5041 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5101 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5161 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5221 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5281 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5341 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5401 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5461 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5521 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5581 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5641 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5701 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5761 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5821 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5881 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5941 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6001 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6061 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6121 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6181 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6241 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6301 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6361 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6421 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6481 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6541 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6601 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6661 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6721 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6781 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6841 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6901 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6961 acgcgaattt t
//
