LOCUS       sequence-193125-        8515 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for N-terminal FLAG-tagged human CCR5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8515)
  AUTHORS   Heredia JD, Park J, Brubaker RJ, Szymanski SK, Gill KS, Procko E
  TITLE     Mapping Interaction Sites on Human Chemokine Receptors by Deep 
            Mutational Scanning.
  JOURNAL   J Immunol. 2018 Apr 20. pii: jimmunol.1800343. doi: 
            10.4049/jimmunol.1800343.
  PUBMED    29678950
REFERENCE   2  (bases 1 to 8515)
  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 Immunol. 
            2018 Apr 20. pii: jimmunol.1800343. doi: 10.4049/jimmunol.1800343."
COMMENT     Sequence Label: pCEP4-FLAG-CCR5
FEATURES             Location/Qualifiers
     source          1..8515
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(576..594)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        662..766
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(767..835,836..1627)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(985..1004)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1798..2386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2287..2306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2817..2962
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             3005..4042
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(4040..4059)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4084..4132
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4786..5165
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5166..5369
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5320..5340
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5366..5390
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5658..5669
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     polyA_signal    6589..6723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6613..6632)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6667..6686
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6866..6888)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcgtcctctt ccccgtcctc gtccatggtt atcaccccct cttctttgag gtccactgcc
       61 gccggagcct tctggtccag atgtgtctcc cttctctcct aggccatttc caggtcctgt
      121 acctggcccc tcgtcagaca tgattcacac taaaagagat caatagacat ctttattaga
      181 cgacgctcag tgaatacagg gagtgcagac tcctgccccc tccaacagcc cccccaccct
      241 catccccttc atggtcgctg tcagacagat ccaggtctga aaattcccca tcctccgaac
      301 catcctcgtc ctcatcacca attactcgca gcccggaaaa ctcccgctga acatcctcaa
      361 gatttgcgtc ctgagcctca agccaggcct caaattcctc gtcccccttt ttgctggacg
      421 gtagggatgg ggattctcgg gacccctcct cttcctcttc aaggtcacca gacagagatg
      481 ctactggggc aacggaagaa aagctgggtg cggcctgtga ggatcagctt atcgatgata
      541 agctgtcaaa catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
      601 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
      661 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      721 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      781 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      841 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      901 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      961 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1021 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1081 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1141 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1201 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1261 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     1321 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1381 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1441 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1501 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1561 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1621 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1681 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1741 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1801 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1861 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1921 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     1981 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2041 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2101 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2161 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2221 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2281 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2341 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2401 ggccttttta cggttcctgg ccttttgctg gccttgaagc tgtccctgat ggtcgtcatc
     2461 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     2521 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     2581 cgcgtcggcc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg
     2641 ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt
     2701 cttggagtgg tgaatccgtt agcgaggtgc cgccctgctt catccccgtg gcccgttgct
     2761 cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga
     2821 cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc
     2881 ggcgcggtcc gaggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc
     2941 gaccctgcag cgacccgctt aacagcgtca acagcgtgcc gcagatcccg gggggcaatg
     3001 agatatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     3061 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     3121 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     3181 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     3241 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     3301 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     3361 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     3421 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     3481 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     3541 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     3601 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     3661 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     3721 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     3781 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     3841 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     3901 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     3961 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccggatcg
     4021 ggagatgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4081 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     4141 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4201 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4261 ctcgcagcca acgtcggggc ggcaggccct gccatagcca ctggccccgt gggttaggga
     4321 cggggtcccc catggggaat ggtttatggt tcgtgggggt tattattttg ggcgttgcgt
     4381 ggggtcaggt ccacgactgg actgagcaga cagacccatg gtttttggat ggcctgggca
     4441 tggaccgcat gtactggcgc gacacgaaca ccgggcgtct gtggctgcca aacacccccg
     4501 acccccaaaa accaccgcgc ggatttctgg cgtgccaagc tagtcgacca attctcatgt
     4561 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     4621 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     4681 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     4741 atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat tgattattga
     4801 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4861 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4921 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4981 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     5041 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     5101 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     5161 ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg
     5221 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     5281 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg
     5341 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc tctagaagct
     5401 gggtaccagc tgctagcgcc accatgaaga ccatcatcgc cctgagctac atcttctgcc
     5461 tggtgttcgc cgactacaag gacgatgacg acggcagccc cggaggcgcg gattatcaag
     5521 tgtcaagtcc aatctatgac atcaattatt atacatcgga gccctgccaa aaaatcaatg
     5581 tgaagcaaat cgcagcccgc ctcctgcctc cgctctactc actggtgttc atctttggtt
     5641 ttgtgggcaa catgctggtc atcctcatcc tgataaactg caaaaggctg aagagcatga
     5701 ctgacatcta cctgctcaac ctggccatct ctgacctgtt tttccttctt actgtcccct
     5761 tctgggctca ctatgctgcc gcccagtggg actttggaaa tacaatgtgt caactcttga
     5821 cagggctcta ttttataggc ttcttctctg gaatcttctt catcatcctc ctgacaatcg
     5881 ataggtacct ggctgtcgtc catgctgtgt ttgctttaaa agccaggacg gtcacctttg
     5941 gggtggtgac aagtgtgatc acttgggtgg tggctgtgtt tgcgtctctc ccaggaatca
     6001 tctttaccag atctcaaaaa gaaggtcttc attacacctg cagctctcat tttccataca
     6061 gtcagtatca attctggaag aatttccaga cattaaagat agtcatcttg gggctggtcc
     6121 tgccgctgct tgtcatggtc atctgctact cgggaatcct aaaaactctg cttcggtgtc
     6181 gaaatgagaa gaagaggcac agggctgtga ggcttatctt caccatcatg attgtttatt
     6241 ttctcttctg ggctccctac aacattgtcc ttctcctgaa caccttccag gaattctttg
     6301 gcctgaataa ttgcagtagc tctaacaggt tggaccaagc tatgcaggtg acagagactc
     6361 ttgggatgac gcactgctgc atcaacccca tcatctatgc ctttgtcggg gagaagttca
     6421 gaaactacct cttagtcttc ttccaaaagc acattgccaa acgcttctgc aaatgctgtt
     6481 ctattttcca gcaagaggct cccgagcgag caagctcagt ttacacccga tccactgggg
     6541 agcaggaaat atctgtgggc ttgtgataac tcgaggccgg caaggccgga tccagacatg
     6601 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     6661 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     6721 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     6781 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc ggctgcctcg
     6841 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     6901 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     6961 gcgggtgtcg gggcgcagcc atgaggtcga ctctagagga tcgatccccg ccccggacga
     7021 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc
     7081 agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca
     7141 aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc
     7201 aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat
     7261 tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc
     7321 ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag
     7381 ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt
     7441 aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct
     7501 aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa
     7561 gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg
     7621 ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca
     7681 aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc
     7741 taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg
     7801 tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta
     7861 tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac
     7921 attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat
     7981 gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca atatgatact
     8041 ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc
     8101 tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc
     8161 taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt
     8221 ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc
     8281 tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc
     8341 actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata
     8401 agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca
     8461 cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcg
//
