LOCUS       sequence-404913-        7025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a mutant form of human RAD23A that lacks the UBA2 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 7025)
  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 7025)
  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 aa 1-317 RAD23A (delta UBA2 do...
FEATURES             Location/Qualifiers
     source          1..7025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     155..177
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     169..186
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     170..186
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        347..726
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        727..930
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     881..901
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     956..975
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        956..974
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1018..1027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1027..1050
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2234..2250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2261..2883
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2460..2479)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2907..2927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2912..3241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3092..3227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3308..4102
                     /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(3362..3381)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3972..3991
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4276..4397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4313..4332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4367..4386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4446..4462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4459..4481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4470..4486
                     /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(4494..4500,4501..4518,4519..4524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4539..4560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4677..4694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4848..5436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4928..4947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5607..6398,6399..6467))
                     /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     6230..6249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6468..6572)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6599..7025,1..2)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6686..6705)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6896..6917
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg
       61 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
      121 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
      181 gccagtgcca agctgatcta tacattgaat caatattggc aattagccat attagtcatt
      241 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
      301 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg
      361 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      421 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      481 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      541 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      601 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      661 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      721 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
      781 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      841 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
      901 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagaa ttttgtaata
      961 cgactcacta tagggcggcc gggaattcgt cgactggatc cggtaccgag gagatctgcc
     1021 accatggatt acaaggatga cgacgataag gcgatcgcca tggagcatat ggccgtcact
     1081 attaccctga agaccctgca gcagcagaca ttcaagattc ggatggaacc tgatgagaca
     1141 gtcaaagtcc tgaaagagaa gatcgaggca gaaaaaggac gagacgcatt ccctgtcgca
     1201 ggacagaagc tgatctacgc tgggaaaatt ctgtccgacg atgtgccaat cagggactat
     1261 cgcattgatg agaagaactt tgtggtcgtg atggtgacta agaccaaagc cggccagggg
     1321 acatctgctc cccctgaagc aagtccaact gccgctcccg agagctccac ctctttccca
     1381 cccgccccaa catccggaat gtctcaccct ccacccgcag cccgcgaaga caaatccccc
     1441 tctgaggaaa gcgcccctac cacatcccca gagagtgtct caggcagcgt gccctctagt
     1501 ggatcaagcg gccgagagga agatgctgca tcaacactgg tgactggaag cgagtacgaa
     1561 accatgctga cagagatcat gagtatgggc tatgagcggg aaagagtcgt ggccgctctg
     1621 cgagccagct acaacaatcc ccaccgggct gtggagtatc tgctgactgg gattcctggc
     1681 agccccgagc ctgaacatgg gtctgtccaa gaaagtcagg tgtcagagca gccagcaacc
     1741 gaggcagcag gagaaaaccc actggagttc ctgagagatc agccccagtt tcagaacatg
     1801 aggcaggtca tccagcagaa tccagcactg ctgcctgctc tgctgcagca gctgggccag
     1861 gaaaaccctc agctgctgca gcagatctcc aggcatcagg agcagtttat tcagatgctg
     1921 aatgaacctc caggggagct ggccgacatc tctgatgtcg agggcgaagt gggggcaatt
     1981 ggagaggaag cccctcagat gaattacatc caggtcacct aacctgtcgc aggagctatt
     2041 gaacggctga aagcactggg cttccccgag agcctggtca tccaggccta ttttgcttgc
     2101 gagaagaacg aaaatctggc tgcaaacttc ctgctgagcc agaattttga cgatgagtaa
     2161 gaattcggat ccacgcgtaa gcttctcgag acgcgttaag cggccgcact cgaggtttaa
     2221 acggccggcc gcggtcatag ctgtttcctg aacagatccc gggtggcatc cctgtgaccc
     2281 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     2341 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     2401 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     2461 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     2521 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     2581 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     2641 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     2701 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca
     2761 gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt
     2821 gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga
     2881 attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2941 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3001 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3061 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3121 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3181 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3241 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3301 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3361 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3421 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3481 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3541 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3601 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3661 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3721 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3781 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3841 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3901 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg ccgaccgcta
     3961 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4021 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4081 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4141 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4201 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4261 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4321 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4381 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4441 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4501 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4561 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4621 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4681 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4741 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4801 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4861 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4921 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4981 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5041 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5101 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5161 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5221 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5281 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5341 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5401 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5461 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5521 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5581 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5641 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5701 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5761 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5821 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5881 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5941 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6001 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6061 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6121 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6181 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6241 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6301 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6361 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6421 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6481 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6541 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6601 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6661 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6721 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6781 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6841 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6901 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     6961 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7021 gcgaa
//
