LOCUS       sequence-395054-        6926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a mutant form of human RAD23B that lacks the N-terminal 
            UBL 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 6926)
  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 6926)
  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_aa 80-409 (delta UBL do...
FEATURES             Location/Qualifiers
     source          1..6926
                     /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(2131..2147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2158..2780
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2357..2376)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2804..2824)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2809..3138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2989..3124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3205..3999
                     /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(3259..3278)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3869..3888
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4173..4294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4210..4229)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4264..4283
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4343..4359)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4356..4378)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4367..4383
                     /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(4391..4397,4398..4415,4416..4421))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4436..4457
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4574..4591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4745..5333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4825..4844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5504..6295,6296..6364))
                     /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     6127..6146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6365..6469)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6496..6924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6583..6602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6793..6814
                     /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 gcatgtgtcc accccagctc
     1081 ccgcaaccac acagcagtct gccccagcta gtactaccgc cgtgacaagc tccacaacta
     1141 ccacagtcgc acaggcacct actccagtgc cagctctggc acccacatca actcctgcca
     1201 gcatcacacc agccagcgct actgcatcta gtgaacctgc cccagcttcc gccgctaagc
     1261 aggagaaacc agctgaaaaa cccgcagaga ctcctgtggc cacctcaccc accgctacag
     1321 acagcacctc cggcgattca agccggtcca acctgttcga ggacgctacc tccgcactgg
     1381 tcacaggcca gtcttacgaa aatatggtga cagagatcat gtctatgggg tatgagcggg
     1441 aacaggtcat tgcagccctg agagctagtt ttaacaatcc cgatagggca gtggaatacc
     1501 tgctgatggg catccctggg gacagagagt ctcaggcagt cgtggacccc cctcaggctg
     1561 caagtaccgg agtcccacag tcctctgcag tggcagctgc agcagctact accacagcta
     1621 ctaccacaac taccagttca ggcgggcacc ctctggagtt cctgcgcaac cagccacagt
     1681 ttcagcagat gcgacagatc attcagcaga atccatccct gctgccagca ctgctgcagc
     1741 agattggacg cgaaaacccc cagctgctgc agcagatctc tcagcaccag gagcatttca
     1801 ttcagatgct gaatgaacct gtgcaggagg caggaggaca gggaggagga ggaggaggcg
     1861 ggagtggagg catcgcagag gccgggtcag gacacatgaa ctacatccag gtcacaccac
     1921 aggagaagga agcaattgaa aggctgaaag ccctgggatt ccccgagggc ctggtcatcc
     1981 aggcctattt tgcttgcgag aagaacgaaa atctggcagc caacttcctg ctgcagcaga
     2041 attttgacga ggattaagaa ttcggatcca cgcgtaagct tctcgagacg cgttaagcgg
     2101 ccgcactcga ggtttaaacg gccggccgcg gtcatagctg tttcctgaac agatcccggg
     2161 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     2221 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     2281 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     2341 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     2401 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     2461 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     2521 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     2581 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     2641 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     2701 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     2761 cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta
     2821 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2881 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2941 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3001 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3061 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3121 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3181 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3241 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3301 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3361 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3421 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3481 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3541 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3601 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3661 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3721 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3781 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3841 ggtgtggccg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3901 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3961 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4021 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4081 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4141 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4201 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4261 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4321 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4381 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4441 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4501 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4561 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4621 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4681 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4741 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4801 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4861 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4921 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4981 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5041 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5101 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5161 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5221 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5281 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5341 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5401 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5461 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5521 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5581 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5641 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5701 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5761 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5821 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5881 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5941 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6001 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6061 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6121 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6181 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6241 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6301 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6361 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6421 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6481 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6541 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6601 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6661 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6721 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6781 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6841 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6901 tttaacaaaa atttaacgcg aatttt
//
