LOCUS       sequence-395043-        6857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a mutant form of human RAD23A that lacks the XPC-binding 
            domain. Variant of plasmid pCMV6-AN-DDK_WT RAD23A..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6857)
  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 6857)
  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 RAD23A_delta aa 230-285 (delta...
FEATURES             Location/Qualifiers
     source          1..6857
                     /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(2062..2078)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2089..2711
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2288..2307)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2735..2755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2740..3069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2920..3055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3136..3930
                     /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(3190..3209)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3800..3819
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4104..4225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4141..4160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4195..4214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4274..4290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4287..4309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4298..4314
                     /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(4322..4328,4329..4346,4347..4352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4367..4388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4505..4522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4676..5264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4756..4775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5435..6226,6227..6295))
                     /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     6058..6077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6296..6400)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6427..6855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6514..6533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6724..6745
                     /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 gcatatggcc gtcactatta
     1081 ccctgaagac cctgcagcag cagacattca agattcggat ggaacctgat gagacagtca
     1141 aagtcctgaa agagaagatc gaggcagaaa aaggacgaga cgcattccct gtcgcaggac
     1201 agaagctgat ctacgctggg aaaattctgt ccgacgatgt gccaatcagg gactatcgca
     1261 ttgatgagaa gaactttgtg gtcgtgatgg tgactaagac caaagccggc caggggacat
     1321 ctgctccccc tgaagcaagt ccaactgccg ctcccgagag ctccacctct ttcccacccg
     1381 ccccaacatc cggaatgtct caccctccac ccgcagcccg cgaagacaaa tccccctctg
     1441 aggaaagcgc ccctaccaca tccccagaga gtgtctcagg cagcgtgccc tctagtggat
     1501 caagcggccg agaggaagat gctgcatcaa cactggtgac tggaagcgag tacgaaacca
     1561 tgctgacaga gatcatgagt atgggctatg agcgggaaag agtcgtggcc gctctgcgag
     1621 ccagctacaa caatccccac cgggctgtgg agtatctgct gactgggatt cctggcagcc
     1681 ccgagcctga acatgggtct gtccaagaaa gtcaggtgtc agagcagcca gcaaccgagg
     1741 cagcaggaga agaacctcca ggggagctgg ccgacatctc tgatgtcgag ggcgaagtgg
     1801 gggcaattgg agaggaagcc cctcagatga attacatcca ggtcacccca caggagaagg
     1861 aagctattga acggctgaaa gcactgggct tccccgagag cctggtcatc caggcctatt
     1921 ttgcttgcga gaagaacgaa aatctggctg caaacttcct gctgagccag aattttgacg
     1981 atgagtaaga attcggatcc acgcgtaagc ttctcgagac gcgttaagcg gccgcactcg
     2041 aggtttaaac ggccggccgc ggtcatagct gtttcctgaa cagatcccgg gtggcatccc
     2101 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     2161 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     2221 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     2281 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     2341 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     2401 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     2461 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     2521 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
     2581 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
     2641 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
     2701 cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga
     2761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2821 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2881 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2941 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3001 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3061 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3121 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3181 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3241 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3301 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3361 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3421 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3481 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3541 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3601 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3661 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3721 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcc
     3781 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3841 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3901 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3961 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4021 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4081 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4141 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4201 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4261 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4321 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4381 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4441 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4501 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4561 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4621 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4681 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4741 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4801 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4861 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4921 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4981 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5041 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5101 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5161 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5221 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5281 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5341 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5401 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5461 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5521 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5581 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5641 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5701 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5761 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5821 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5881 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5941 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6001 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6061 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6121 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6181 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6241 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6301 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6361 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6421 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     6481 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     6541 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     6601 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     6661 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6721 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     6781 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6841 aatttaacgc gaatttt
//
