LOCUS       sequence-181230-        6412 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression plasmid for untagged human CCR5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6412)
  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 6412)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pcDNA3-CCR5
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1057..1068
                     /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)"
     primer_bind     complement(2006..2023)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2012..2236
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2282..2710
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2369..2388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2579..2600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2719..2739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2724..3053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2904..3039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2966..2985
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3120..3914
                     /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(3174..3193)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3784..3803
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4088..4209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4125..4144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4179..4198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4258..4274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4258..4274)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4271..4293)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4282..4298
                     /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(4306..4312,4313..4330,4331..4336))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4351..4372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4489..4506)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4660..5245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4740..4759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5416..6207,6208..6276))
                     /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     6039..6058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6277..6381)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg attatcaagt gtcaagtcca atctatgaca tcaattatta tacatcggag
      961 ccctgccaaa aaatcaatgt gaagcaaatc gcagcccgcc tcctgcctcc gctctactca
     1021 ctggtgttca tctttggttt tgtgggcaac atgctggtca tcctcatcct gataaactgc
     1081 aaaaggctga agagcatgac tgacatctac ctgctcaacc tggccatctc tgacctgttt
     1141 ttccttctta ctgtcccctt ctgggctcac tatgctgccg cccagtggga ctttggaaat
     1201 acaatgtgtc aactcttgac agggctctat tttataggct tcttctctgg aatcttcttc
     1261 atcatcctcc tgacaatcga taggtacctg gctgtcgtcc atgctgtgtt tgctttaaaa
     1321 gccaggacgg tcacctttgg ggtggtgaca agtgtgatca cttgggtggt ggctgtgttt
     1381 gcgtctctcc caggaatcat ctttaccaga tctcaaaaag aaggtcttca ttacacctgc
     1441 agctctcatt ttccatacag tcagtatcaa ttctggaaga atttccagac attaaagata
     1501 gtcatcttgg ggctggtcct gccgctgctt gtcatggtca tctgctactc gggaatccta
     1561 aaaactctgc ttcggtgtcg aaatgagaag aagaggcaca gggctgtgag gcttatcttc
     1621 accatcatga ttgtttattt tctcttctgg gctccctaca acattgtcct tctcctgaac
     1681 accttccagg aattctttgg cctgaataat tgcagtagct ctaacaggtt ggaccaagct
     1741 atgcaggtga cagagactct tgggatgacg cactgctgca tcaaccccat catctatgcc
     1801 tttgtcgggg agaagttcag aaactacctc ttagtcttct tccaaaagca cattgccaaa
     1861 cgcttctgca aatgctgttc tattttccag caagaggctc ccgagcgagc aagctcagtt
     1921 tacacccgat ccactgggga gcaggaaata tctgtgggct tgtgataact cgagtctaga
     1981 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2041 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2101 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2161 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2221 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2281 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2341 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2401 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2461 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2521 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2581 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2641 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2701 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2761 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2881 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2941 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3001 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3061 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3121 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3181 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3241 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3301 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3361 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3421 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3481 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3541 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3601 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3661 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3721 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3781 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3841 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3901 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     3961 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4021 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4081 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4141 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4201 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4261 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4321 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4381 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4441 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4501 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4561 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4621 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4681 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4741 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4801 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4861 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4921 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4981 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5041 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5101 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5161 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     5221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5281 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5341 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5401 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5461 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5521 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5581 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5641 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5701 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5761 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5821 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5881 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5941 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6001 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6061 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6121 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6181 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6241 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6301 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6361 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
