LOCUS       sequence-404909-        6695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a mutant form of human RAD23A that lacks the N-terminal 
            UBL 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 6695)
  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 6695)
  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_aa 111-363 (delta UBL d...
FEATURES             Location/Qualifiers
     source          1..6695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     194..211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     195..211
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        372..751
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        752..955
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     906..926
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     981..1000
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        981..999
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1043..1052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1052..1075
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1929..1945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1956..2578
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2155..2174)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2602..2622)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2607..2936
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2787..2922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3003..3797
                     /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(3057..3076)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3667..3686
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3971..4092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4008..4027)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4062..4081
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4141..4157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4154..4176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4165..4181
                     /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(4189..4195,4196..4213,4214..4219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4234..4255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4372..4389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4543..5131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4623..4642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5302..6093,6094..6162))
                     /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     5925..5944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6163..6267)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6294..6695,1..54)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6381..6400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6591..6612
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc
       61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      181 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg atctatacat tgaatcaata
      241 ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg gctattggcc
      301 attgcatacg ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg
      361 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      421 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      481 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      541 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      601 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
      661 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
      721 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
      781 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      841 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
      901 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
      961 agtgaaccgt cagaattttg taatacgact cactataggg cggccgggaa ttcgtcgact
     1021 ggatccggta ccgaggagat ctgccaccat ggattacaag gatgacgacg ataaggcgat
     1081 cgccatggag catatgtctc accctccacc cgcagcccgc gaagacaaat ccccctctga
     1141 ggaaagcgcc cctaccacat ccccagagag tgtctcaggc agcgtgccct ctagtggatc
     1201 aagcggccga gaggaagatg ctgcatcaac actggtgact ggaagcgagt acgaaaccat
     1261 gctgacagag atcatgagta tgggctatga gcgggaaaga gtcgtggccg ctctgcgagc
     1321 cagctacaac aatccccacc gggctgtgga gtatctgctg actgggattc ctggcagccc
     1381 cgagcctgaa catgggtctg tccaagaaag tcaggtgtca gagcagccag caaccgaggc
     1441 agcaggagaa aacccactgg agttcctgag agatcagccc cagtttcaga acatgaggca
     1501 ggtcatccag cagaatccag cactgctgcc tgctctgctg cagcagctgg gccaggaaaa
     1561 ccctcagctg ctgcagcaga tctccaggca tcaggagcag tttattcaga tgctgaatga
     1621 acctccaggg gagctggccg acatctctga tgtcgagggc gaagtggggg caattggaga
     1681 ggaagcccct cagatgaatt acatccaggt caccccacag gagaaggaag ctattgaacg
     1741 gctgaaagca ctgggcttcc ccgagagcct ggtcatccag gcctattttg cttgcgagaa
     1801 gaacgaaaat ctggctgcaa acttcctgct gagccagaat tttgacgatg agtaagaatt
     1861 cggatccacg cgtaagcttc tcgagacgcg ttaagcggcc gcactcgagg tttaaacggc
     1921 cggccgcggt catagctgtt tcctgaacag atcccgggtg gcatccctgt gacccctccc
     1981 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     2041 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     2101 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     2161 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     2221 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     2281 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     2341 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     2401 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     2461 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     2521 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     2581 gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2641 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2701 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2761 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2821 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2881 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2941 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3001 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3061 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3121 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3181 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3241 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3301 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3361 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3421 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3481 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3541 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3601 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggccgac cgctatcagg
     3661 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3721 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3781 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3841 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3901 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3961 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4021 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4081 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4141 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4201 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4261 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4321 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4381 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4441 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4501 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4561 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4621 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4681 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4741 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4801 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4861 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4921 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4981 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5041 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5101 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5161 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5221 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5281 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5341 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5401 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5461 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5521 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5581 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5641 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5701 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5761 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5821 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5881 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5941 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6001 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6061 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6121 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6181 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6241 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     6301 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6361 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6421 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6481 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6541 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6601 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6661 tttgccgatt tcggcctatt ggttaaaaaa tgagc
//
