LOCUS       sequence-423351-        6963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RPA2. N-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6963)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 6963)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pHTN-HaloTag_RPA2:1-270
FEATURES             Location/Qualifiers
     source          1..6963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(2914..2932)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2964..3085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3029..3048
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3330..3687
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3330..3521
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3353..3373)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3538..3673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3738..4532
                     /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(3792..3811)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4402..4421
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4596..4644
                     /note="synthetic polyadenylation signal"
     promoter        4800..4904
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4905..4973,4974..5765)
                     /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     complement(5123..5142)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5923..6511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6003..6022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6627..6910
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgcttccga attcagagct caaccgcgga tatctatgtg
     2041 gaacagtgga ttcgaaagct atggcagctc ctcatacggg ggagccggcg gctacacgca
     2101 gtccccgggg ggctttggat cgcccgcacc ttctcaagcc gaaaagaaat caagagcccg
     2161 agcccagcac attgtgccct gtactatatc tcagctgctt tctgccactt tggttgatga
     2221 agtgttcaga attgggaatg ttgagatttc acaggtcact attgtgggga tcatcagaca
     2281 tgcagagaag gctccaacca acattgttta caaaatagat gacatgacag ctgcacccat
     2341 ggacgttcgc cagtgggttg acacagatga caccagcagt gaaaacactg tggttcctcc
     2401 agaaacatat gtgaaagtgg caggccacct gagatctttt cagaacaaaa agagcctggt
     2461 agcctttaag atcatgcccc tggaggatat gaatgagttc accacacata ttctggaagt
     2521 gatcaatgca cacatggtac taagcaaagc caacagccag ccctcagcag ggagagcacc
     2581 tatcagcaat ccaggaatga gtgaagcagg gaactttggt gggaatagct tcatgccagc
     2641 aaatggcctc actgtggccc aaaaccaggt gttgaatttg attaaggctt gtccaagacc
     2701 tgaagggttg aactttcagg atctcaagaa ccagctgaaa cacatgtctg tatcctcaat
     2761 caagcaagct gtggattttc tgagcaatga ggggcacatc tattctactg tggatgatga
     2821 ccattttaaa tccacagatg cagaataaga tatctagatt tgggcccaat tcctgcagga
     2881 ttttgcggcc gcttgctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     2941 ttggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3001 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3061 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3121 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3181 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     3241 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     3301 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     3361 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3421 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3481 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3601 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3661 ttttttggag gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac
     3721 ttaaggctag agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3781 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3841 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3901 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3961 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4021 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4081 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4141 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4201 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4261 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4321 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4381 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4441 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4501 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4561 ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac
     4621 atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt
     4681 tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg
     4741 gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4801 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4861 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4921 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4981 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5041 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5101 tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5161 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5221 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5281 ccatgagtga taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc
     5341 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5401 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5461 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5521 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5581 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5641 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5701 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5761 ggtaattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     5821 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5881 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5941 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6001 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6061 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6121 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6181 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6241 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6301 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6361 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6421 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6481 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     6541 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6601 gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt
     6661 cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg
     6721 gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg
     6781 gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac
     6841 gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg
     6901 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
     6961 tct
//
