LOCUS       sequence-395061-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a variant of human RAD23B containing mutation V222A within
            UBA1. Variant of plasmid pCMV6-AN-DDK_WT RAD23B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  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 7163)
  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_V222A RAD23B
FEATURES             Location/Qualifiers
     source          1..7163
                     /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(2368..2384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2395..3017
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2594..2613)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3041..3061)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3046..3375
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3226..3361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3442..4236
                     /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(3496..3515)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4106..4125
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4410..4531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4447..4466)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4501..4520
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4580..4596)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4593..4615)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4604..4620
                     /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(4628..4634,4635..4652,4653..4658))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4673..4694
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4811..4828)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4982..5570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5062..5081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5741..6532,6533..6601))
                     /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     6364..6383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6602..6706)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6733..7161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6820..6839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7030..7051
                     /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 tagggcagcc gagtacttgc
     1741 tgatgggcat ccctggggac agagagtctc aggcagtcgt ggacccccct caggctgcaa
     1801 gtaccggagt cccacagtcc tctgcagtgg cagctgcagc agctactacc acagctacta
     1861 ccacaactac cagttcaggc gggcaccctc tggagttcct gcgcaaccag ccacagtttc
     1921 agcagatgcg acagatcatt cagcagaatc catccctgct gccagcactg ctgcagcaga
     1981 ttggacgcga aaacccccag ctgctgcagc agatctctca gcaccaggag catttcattc
     2041 agatgctgaa tgaacctgtg caggaggcag gaggacaggg aggaggagga ggaggcggga
     2101 gtggaggcat cgcagaggcc gggtcaggac acatgaacta catccaggtc acaccacagg
     2161 agaaggaagc aattgaaagg ctgaaagccc tgggattccc cgagggcctg gtcatccagg
     2221 cctattttgc ttgcgagaag aacgaaaatc tggcagccaa cttcctgctg cagcagaatt
     2281 ttgacgagga ttaagaattc ggatccacgc gtaagcttct cgagacgcgt taagcggccg
     2341 cactcgaggt ttaaacggcc ggccgcggtc atagctgttt cctgaacaga tcccgggtgg
     2401 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     2461 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     2521 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     2581 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     2641 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     2701 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     2761 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     2821 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta
     2881 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa
     2941 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag
     3001 tagatgcctg ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg
     3061 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3121 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3181 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3241 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3301 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3361 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3421 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3481 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3541 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3601 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3661 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3721 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3781 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3841 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3901 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3961 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4021 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4081 gtggccgacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4141 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4201 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4261 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4321 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4381 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4441 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4501 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4561 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4621 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4681 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4741 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4801 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4861 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4921 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4981 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5041 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5101 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5161 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5221 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5281 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5341 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5401 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5461 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5521 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5581 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5641 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5701 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5761 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5821 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5881 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5941 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6001 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6061 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6121 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6181 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6241 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6301 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6361 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6421 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6481 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6541 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6601 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6661 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6721 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6781 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6841 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6901 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6961 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     7021 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     7081 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     7141 aacaaaaatt taacgcgaat ttt
//
