LOCUS       sequence-395053-        7040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a mutant form of human RAD23B that lacks the UBA1 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 7040)
  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 7040)
  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 188-228 (delta...
FEATURES             Location/Qualifiers
     source          1..7040
                     /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(2245..2261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2272..2894
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2471..2490)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2918..2938)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2923..3252
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3103..3238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3319..4113
                     /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(3373..3392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3983..4002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4287..4408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4324..4343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4378..4397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4457..4473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4470..4492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4481..4497
                     /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(4505..4511,4512..4529,4530..4535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4550..4571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4688..4705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4859..5447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4939..4958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5618..6409,6410..6478))
                     /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     6241..6260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6479..6583)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6610..7038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6697..6716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6907..6928
                     /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 caggcatccc tggggacaga gagtctcagg cagtcgtgga cccccctcag gctgcaagta
     1681 ccggagtccc acagtcctct gcagtggcag ctgcagcagc tactaccaca gctactacca
     1741 caactaccag ttcaggcggg caccctctgg agttcctgcg caaccagcca cagtttcagc
     1801 agatgcgaca gatcattcag cagaatccat ccctgctgcc agcactgctg cagcagattg
     1861 gacgcgaaaa cccccagctg ctgcagcaga tctctcagca ccaggagcat ttcattcaga
     1921 tgctgaatga acctgtgcag gaggcaggag gacagggagg aggaggagga ggcgggagtg
     1981 gaggcatcgc agaggccggg tcaggacaca tgaactacat ccaggtcaca ccacaggaga
     2041 aggaagcaat tgaaaggctg aaagccctgg gattccccga gggcctggtc atccaggcct
     2101 attttgcttg cgagaagaac gaaaatctgg cagccaactt cctgctgcag cagaattttg
     2161 acgaggatta agaattcgga tccacgcgta agcttctcga gacgcgttaa gcggccgcac
     2221 tcgaggttta aacggccggc cgcggtcata gctgtttcct gaacagatcc cgggtggcat
     2281 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     2341 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     2401 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     2461 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     2521 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     2581 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     2641 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     2701 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa
     2761 taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg
     2821 gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag
     2881 atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt
     2941 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3001 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3061 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3121 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3181 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3241 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3301 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3361 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3421 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3481 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3541 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3601 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3661 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3721 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3781 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3841 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3901 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3961 gccgaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4021 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4081 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4141 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4201 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4261 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4321 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4381 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4441 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4501 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4561 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4621 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6601 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6661 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6721 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6781 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6841 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6901 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6961 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     7021 aaaaatttaa cgcgaatttt
//
