LOCUS       sequence-394878-        7166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human RAD23B with an N-terminal HA tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7166)
  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 7166)
  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-HA_WT RAD23B
FEATURES             Location/Qualifiers
     source          1..7166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     140..162
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     154..171
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     155..171
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        332..711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        712..915
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     866..886
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     941..960
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        941..959
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1012..1038
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2360..2376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2387..3009
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2586..2605)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3033..3053)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3038..3367
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3218..3353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3434..4228
                     /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(3488..3507)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4098..4117
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4402..4523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4439..4458)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4493..4512
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4572..4588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4585..4607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4596..4612
                     /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(4620..4626,4627..4644,4645..4650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4665..4686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4803..4820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4974..5562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5054..5073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5733..6524,6525..6593))
                     /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     6356..6375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6594..6698)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6725..7153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6812..6831)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7022..7043
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
       61 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
      121 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
      181 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
      241 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      301 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
      361 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      421 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      481 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      541 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      601 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      661 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      721 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      781 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      841 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      901 tatataagca gagctcgttt agtgaaccgt cagaattttg taatacgact cactataggg
      961 cggccgggaa ttcgtcgact ggatccggta ccgaggagat ctgccaccat gtacccatac
     1021 gatgttccag attacgctgc gatcgccatg gagcatatgc aggtgacact gaagacactg
     1081 cagcagcaga cttttaagat tgacattgac cccgaggaga ccgtgaaggc actgaaggag
     1141 aaaatcgaga gcgaaaaggg caaagacgca ttccctgtgg ccgggcagaa gctgatctac
     1201 gccggaaaaa ttctgaacga cgataccgct ctgaaggaat acaagatcga tgagaagaat
     1261 ttcgtggtcg tgatggtcac taagcctaaa gccgtgtcca ccccagctcc cgcaaccaca
     1321 cagcagtctg ccccagctag tactaccgcc gtgacaagct ccacaactac cacagtcgca
     1381 caggcaccta ctccagtgcc agctctggca cccacatcaa ctcctgccag catcacacca
     1441 gccagcgcta ctgcatctag tgaacctgcc ccagcttccg ccgctaagca ggagaaacca
     1501 gctgaaaaac ccgcagagac tcctgtggcc acctcaccca ccgctacaga cagcacctcc
     1561 ggcgattcaa gccggtccaa cctgttcgag gacgctacct ccgcactggt cacaggccag
     1621 tcttacgaaa atatggtgac agagatcatg tctatggggt atgagcggga acaggtcatt
     1681 gcagccctga gagctagttt taacaatccc gatagggcag tggaatacct gctgatgggc
     1741 atccctgggg acagagagtc tcaggcagtc gtggaccccc ctcaggctgc aagtaccgga
     1801 gtcccacagt cctctgcagt ggcagctgca gcagctacta ccacagctac taccacaact
     1861 accagttcag gcgggcaccc tctggagttc ctgcgcaacc agccacagtt tcagcagatg
     1921 cgacagatca ttcagcagaa tccatccctg ctgccagcac tgctgcagca gattggacgc
     1981 gaaaaccccc agctgctgca gcagatctct cagcaccagg agcatttcat tcagatgctg
     2041 aatgaacctg tgcaggaggc aggaggacag ggaggaggag gaggaggcgg gagtggaggc
     2101 atcgcagagg ccgggtcagg acacatgaac tacatccagg tcacaccaca ggagaaggaa
     2161 gcaattgaaa ggctgaaagc cctgggattc cccgagggcc tggtcatcca ggcctatttt
     2221 gcttgcgaga agaacgaaaa tctggcagcc aacttcctgc tgcagcagaa ttttgacgag
     2281 gattaagaat tcggatccac gcgtaagctt ctcgagacgc gttaagcggc cgcactcgag
     2341 gtttaaacgg ccggccgcgg tcatagctgt ttcctgaaca gatcccgggt ggcatccctg
     2401 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     2461 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     2521 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     2581 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     2641 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     2701 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     2761 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     2821 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     2881 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     2941 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3001 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3061 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3121 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3181 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3241 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3301 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3361 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3421 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3481 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3541 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3601 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3661 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3721 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3781 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3841 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3901 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3961 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4021 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggccga
     4081 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4141 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4201 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4261 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4321 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4381 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4441 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4501 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4561 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4621 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4681 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4741 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5401 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6601 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6661 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6721 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6781 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6841 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6901 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6961 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7021 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7081 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7141 tttaacgcga attttaacaa aatatt
//
