LOCUS       sequence-331353-        6667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in HEK293T cell and compete ligand singaling against 
            full-length receptors, the truncated receptor can perform signaling 
            at high ligand concentration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6667)
  AUTHORS   Qing R, Tao F, Chatterjee P, Yang G, Han Q, Chung H, Ni J, Suter BP,
            Kubicek J, Maertens B, Schubert T, Blackburn C, Zhang S
  TITLE     Non-full-length Water-Soluble CXCR4(QTY) and CCR5(QTY) Chemokine 
            Receptors: Implication for Overlooked Truncated but Functional 
            Membrane Receptors.
  JOURNAL   iScience. 2020 Oct 28;23(12):101670. doi: 
            10.1016/j.isci.2020.101670. eCollection 2020 Dec 18.
  PUBMED    33376963
REFERENCE   2  (bases 1 to 6667)
  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: "iScience.";
            date: "2020-10-28"; volume: "23(12):101670. doi"; pages: " 
            10.1016/j.isci.2020.101670. eCollection 2020 Dec 18"
COMMENT     Sequence Label: CCR5-SZ190b-sfGFP
FEATURES             Location/Qualifiers
     source          1..6667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..65
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             200..994
                     /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(254..273)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     864..883
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1168..1289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1205..1224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1259..1278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1338..1354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1338..1354)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1351..1373)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1362..1378
                     /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(1386..1392,1393..1410,1411..1416))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1431..1452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1569..1586)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1740..2328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1820..1839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2499..3290,3291..3359))
                     /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     3122..3141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3360..3464)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3539..3558)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3730..4109
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4110..4313
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4264..4284
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4358..4377
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4358..4376
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4396..4405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4552..4563
                     /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)"
     CDS             4999..5712
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4999..5709
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(5026..5054)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5649..5669
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5753..5770)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5759..5983
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6029..6457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6116..6135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6326..6347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6466..6486)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6471..6667,1..133)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6651..6667,1..119)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
       61 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
      121 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
      181 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
      241 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
      301 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
      361 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
      421 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
      481 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
      541 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
      601 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
      661 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
      721 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
      781 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
      841 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
      901 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
      961 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     1021 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     1081 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     1141 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     1201 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     1261 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     1321 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1381 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1441 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1501 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1561 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1621 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1681 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1741 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1801 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1861 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1921 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1981 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2041 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2101 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2161 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2221 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2281 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2341 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2401 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2461 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2521 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2581 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2641 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2701 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2761 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2821 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2881 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2941 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3001 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3061 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3121 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3181 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3241 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3301 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3361 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3421 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3481 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     3541 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     3601 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     3661 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     3721 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     3781 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3841 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3901 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3961 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4021 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4081 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     4141 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4201 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4261 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     4321 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     4381 cccaagctgg ctagcgccac catggattat caagtgtcaa gtccaatcta tgacatcaat
     4441 tattatacat cggagccctg ccaaaaaatc aatgtgaagc aaatcgcagc ccgcctcctg
     4501 cctccgctct actcactggt gttcatcttt ggttttgtgg gcaacatgct ggtcatcctc
     4561 atcctgataa actgcaaaag gctgaagagc atgactgaca tctacctgct caacctggcc
     4621 atctctgacc tgtttttcct tcttactgtc cccttctggg ctccctacaa cattgtcctt
     4681 ctcctgaaca ccttccagga attctttggc ctgaataatt gcagtagctc taacaggttg
     4741 gaccaagcta tgcaggtgac agagactctt gggatgacgc actgctgcat caaccccatc
     4801 atctatgcct ttgtcgggga gaagttcaga aactacctct tagtcttctt ccaaaagcac
     4861 attgccaaac gcttctgcaa atgctgttct attttccagc aagaggctcc cgagcgagca
     4921 agctcagttt acacccgatc cactggggag caggaaatat ctgtgggctt gggatccggt
     4981 gggtccggta gtggtggttc caaaggagaa gaacttttca ctggagttgt cccaattctt
     5041 gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt
     5101 gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt
     5161 ccatggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg
     5221 gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
     5281 cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa
     5341 ggtgataccc ttgttaatcg tatcgagtta aaaggtattg attttaaaga agatggaaac
     5401 attctcggac acaaactgga gtacaacttt aactcacaca atgtatacat cacggcagac
     5461 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc
     5521 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     5581 ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa cgaaaagcgt
     5641 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
     5701 ctctacaaat gataactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     5761 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5821 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5881 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5941 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     6001 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     6061 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     6121 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     6181 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     6241 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     6301 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     6361 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     6421 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     6481 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6541 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6601 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     6661 taactcc
//
