LOCUS       sequence-423350-        6473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length HV1N5 VPR. C-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6473)
  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 6473)
  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: pHTC-HaloTag_VPR:1-96
FEATURES             Location/Qualifiers
     source          1..6473
                     /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      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1427..1447
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1463..2347
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(2419..2437)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2474..2595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2485..2504)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2539..2558
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2840..3197
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2840..3031
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2863..2883)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3048..3183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3248..4042
                     /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(3302..3321)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3912..3931
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4106..4154
                     /note="synthetic polyadenylation signal"
     promoter        4310..4414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4415..4483,4484..5275)
                     /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(4633..4652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5433..6021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5513..5532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6137..6420
                     /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 gcgatcgctt ccgaattcgc
     1081 caccatggag caggcccccg aggaccaggg cccccagcgc gagccctaca acgagtggac
     1141 cctggagctg ctggaggagc tgaagagcga ggccgtgcgc cacttccccc gcatctggct
     1201 gcacaacctg ggccagcaca tctacgagac ctacggcgac acctgggccg gcgtggaggc
     1261 catcatccgc atcctgcagc agctgctgtt catccacttc cgcatcggct gccgccacag
     1321 ccgcatcggc gtgacccgcc agcgccgcgc ccgcaacggc gccagccgca gccggatatc
     1381 tagatttggg cccaattcct gcaggcgagc tctcgagcca accactgagg atctgtactt
     1441 tcagagcgat aacgatggat ccgaaatcgg tactggcttt ccattcgacc cccattatgt
     1501 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt
     1561 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt
     1621 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc
     1681 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct
     1741 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg
     1801 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat
     1861 cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac
     1921 cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat
     1981 gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa
     2041 tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc
     2101 agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc
     2161 gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct
     2221 ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca
     2281 agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcta ctctggagat
     2341 ttccggttaa tagaattggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag
     2401 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt
     2461 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2521 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2581 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2641 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa
     2701 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     2761 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
     2821 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     2881 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2941 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3001 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3061 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3121 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3181 gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac ttaaggctag
     3241 agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3301 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3361 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3421 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3481 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3541 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3601 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3661 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3721 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3781 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3841 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3901 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3961 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4021 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4081 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4141 tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt
     4201 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     4261 ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4321 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4381 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4441 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4501 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4561 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4621 tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4681 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4741 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4801 taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc taaccgcttt
     4861 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4921 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4981 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5041 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5101 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5161 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5221 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaattcga
     5281 aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta
     5341 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5401 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5461 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5521 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5581 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5641 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5701 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5761 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5821 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5881 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5941 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6001 cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6061 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6121 tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg
     6181 gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc
     6241 tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc
     6301 gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc
     6361 ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga
     6421 gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
