LOCUS       sequence-193149-        9299 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  FLAG-tagged human CXCR4 fused to N-terminus of split Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9299)
  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 9299)
  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-CXCR4-VN
FEATURES             Location/Qualifiers
     source          1..9299
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(716..734)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        802..906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(907..975,976..1767)
                     /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(1125..1144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1938..2526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2427..2446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2957..3102
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             3145..4182
                     /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(4180..4199)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4224..4272
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4926..5305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5306..5509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5460..5480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5506..5530
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6371..6382
                     /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             join(6704..6706,6707..7165)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYLTA"
     primer_bind     complement(6746..6767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    7188..7322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7212..7231)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7266..7285
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7465..7487)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cctgctcctg ctcctgttcc accgtgggtc cctttgcagc caatgcaact tggacgtttt
       61 tggggtctcc ggacaccatc tctatgtctt ggccctgatc ctgagccgcc cggggctcct
      121 ggtcttccgc ctcctcgtcc tcgtcctctt ccccgtcctc gtccatggtt atcaccccct
      181 cttctttgag gtccactgcc gccggagcct tctggtccag atgtgtctcc cttctctcct
      241 aggccatttc caggtcctgt acctggcccc tcgtcagaca tgattcacac taaaagagat
      301 caatagacat ctttattaga cgacgctcag tgaatacagg gagtgcagac tcctgccccc
      361 tccaacagcc cccccaccct catccccttc atggtcgctg tcagacagat ccaggtctga
      421 aaattcccca tcctccgaac catcctcgtc ctcatcacca attactcgca gcccggaaaa
      481 ctcccgctga acatcctcaa gatttgcgtc ctgagcctca agccaggcct caaattcctc
      541 gtcccccttt ttgctggacg gtagggatgg ggattctcgg gacccctcct cttcctcttc
      601 aaggtcacca gacagagatg ctactggggc aacggaagaa aagctgggtg cggcctgtga
      661 ggatcagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc
      721 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
      781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      841 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      901 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      961 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1021 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1081 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1141 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1201 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1261 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1321 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1381 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1441 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1501 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1561 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1621 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1681 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1741 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1801 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1861 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1921 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1981 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2041 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2101 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2161 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2221 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2281 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2341 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2401 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2461 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2521 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttgaagc
     2581 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
     2641 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
     2701 gccagcaaga cgtagcccag cgcgtcggcc ccgagatgcg ccgcgtgcgg ctgctggaga
     2761 tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca
     2821 agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccctgctt
     2881 catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca
     2941 tgtctttagt tctatgatga cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa
     3001 cacgcagatg cagtcggggc ggcgcggtcc gaggtccact tcgcatatta aggtgacgcg
     3061 tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aacagcgtca acagcgtgcc
     3121 gcagatcccg gggggcaatg agatatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     3181 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     3241 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     3301 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     3361 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     3421 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     3481 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     3541 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     3601 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     3661 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     3721 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     3781 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     3841 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     3901 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     3961 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     4021 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     4081 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     4141 cccagcactc gtccggatcg ggagatgggg gaggctaact gaaacacgga aggagacaat
     4201 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     4261 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4321 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4381 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcca
     4441 ctggccccgt gggttaggga cggggtcccc catggggaat ggtttatggt tcgtgggggt
     4501 tattattttg ggcgttgcgt ggggtcaggt ccacgactgg actgagcaga cagacccatg
     4561 gtttttggat ggcctgggca tggaccgcat gtactggcgc gacacgaaca ccgggcgtct
     4621 gtggctgcca aacacccccg acccccaaaa accaccgcgc ggatttctgg cgtgccaagc
     4681 tagtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa cgttgttgcc
     4741 attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc aatattggca
     4801 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     4861 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     4921 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     4981 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5041 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5101 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5161 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     5221 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     5281 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     5341 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     5401 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     5461 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     5521 ccgtcagatc tctagaagct gggtaccagc tgctagcgcc accatgaaga ccatcatcgc
     5581 cctgagctac atcttctgcc tggtgttcgc cgactacaag gatgacgacg acggcagccc
     5641 cggaggagct gaggggatca gtatatacac ttcagataac tacaccgagg aaatgggctc
     5701 aggggactat gactccatga aggaaccctg tttccgtgaa gaaaatgcta atttcaataa
     5761 aatcttcctg cccaccatct actccatcat cttcttaact ggcattgtgg gcaatggatt
     5821 ggtcatcctg gtcatgggtt accagaagaa actgagaagc atgacggaca agtacaggct
     5881 gcacctgtca gtggccgacc tcctctttgt catcacgctt cccttctggg cagttgatgc
     5941 cgtggcaaac tggtactttg ggaacttcct atgcaaggca gtccatgtca tctacacagt
     6001 caacctctac agcagtgtcc tcatcctggc cttcatcagt ctggaccgct acctggccat
     6061 cgtccacgcc accaacagtc agaggccaag gaagctgttg gctgaaaagg tggtctatgt
     6121 tggcgtctgg atccctgccc tcctgctgac tattcccgac ttcatctttg ccaacgtcag
     6181 tgaggcagat gacagatata tctgtgaccg cttctacccc aatgacttgt gggtggttgt
     6241 gttccagttt cagcacatca tggttggcct tatcctgcct ggtattgtca tcctgtcctg
     6301 ctattgcatt atcatctcca agctgtcaca ctccaagggc caccagaagc gcaaggccct
     6361 caagaccaca gtcatcctca tcctggcttt cttcgcctgt tggctgcctt actacattgg
     6421 gatcagcatc gactccttca tcctcctgga aatcatcaag caagggtgtg agtttgagaa
     6481 cactgtgcac aagtggattt ccatcaccga ggccctagct ttcttccact gttgtctgaa
     6541 ccccatcctc tatgctttcc ttggagccaa atttaaaacc tctgcccagc acgcactcac
     6601 ctctgtgagc agagggtcca gcctcaagat cctctccaaa ggaaagcgag gtggacattc
     6661 atctgtttcc actgagtctg agtcttcaag ttttcactcc agcgtgagca agggcgagga
     6721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     6781 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     6841 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     6901 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     6961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     7021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     7081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     7141 cagccacaac gtctatctga ccgcctaact cgaggccggc aaggccggat ccagacatga
     7201 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     7261 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     7321 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     7381 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatccg gctgcctcgc
     7441 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     7501 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     7561 cgggtgtcgg ggcgcagcca tgaggtcgac tctagaggat cgatccccgc cccggacgaa
     7621 ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt aatcccttca
     7681 gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg gggggaccaa
     7741 acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc acatttgcca
     7801 acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca acacaacatt
     7861 gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac atgtacctcc
     7921 caggggccca ggaagactac gggaggctac accaacgtca atcagagggg cctgtgtagc
     7981 taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc ccccttgtta
     8041 accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag actaacccta
     8101 attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg ttagtaaaag
     8161 ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta tttacatggg
     8221 gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa
     8281 ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct tcgtttagct
     8341 aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt
     8401 ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc attgtgctat
     8461 gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag gaatgaaaca
     8521 ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa agactggatg
     8581 tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa tatgatactg
     8641 gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt gttacactct
     8701 atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact ggttgtctct
     8761 aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca aagacaagtg
     8821 gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac acgccgccct
     8881 gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc ccgctaacca
     8941 ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat acctgcataa
     9001 gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca aattacacac
     9061 acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag gtcgctgaga
     9121 gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg gatagcatat
     9181 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     9241 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcata
//
