LOCUS       sequence-225501-        7488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CD4 fused to TagRFP-T.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7488)
  AUTHORS   Pereira PM, Albrecht D, Culley S, Jacobs C, Marsh M, Mercer J, 
            Henriques R
  TITLE     Fix Your Membrane Receptor Imaging: Actin Cytoskeleton and CD4 
            Membrane Organization Disruption by Chemical Fixation.
  JOURNAL   Front Immunol. 2019 Apr 5;10:675. doi: 10.3389/fimmu.2019.00675. 
            eCollection 2019.
  PUBMED    31024536
REFERENCE   2  (bases 1 to 7488)
  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: "Front 
            Immunol."; date: "2019-04-5"; volume: "10:675. doi"; pages: " 
            10.3389/fimmu.2019.00675. eCollection 2019"
COMMENT     Sequence Label: pCD4-TagRFP-T (RHPMP151)
FEATURES             Location/Qualifiers
     source          1..7488
                     /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"
     CDS             join(927..1001,1002..2300)
                     /codon_start=1
                     /product="human CD4 cell surface glycoprotein"
                     /label=CD4
                     /translation="MNRGVPFRHLLLVLQLALLPAATQGKKVVLGKKGDTVELTCTASQ
                     KKSIQFHWKNSNQIKILGNQGSFLTKGPSKLNDRADSRRSLWDQGNFPLIIKNLKIEDS
                     DTYICEVEDQKEEVQLLVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRGKNI
                     QGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFQKASSIVYKKEGEQVEFS
                     FPLAFTVEKLTGSGELWWQAERASSSKSWITFDLKNKEVSVKRVTQDPKLQMGKKLPLH
                     LTLPQALPQYAGSGNLTLALEAKTGKLHQEVNLVVMRATQLQKNLTCEVWGPTSPKLML
                     SLKLENKEAKVSKREKAVWVLNPEAGMWQCLLSDSGQVLLESNIKVLPTWSTPVQPMAL
                     IVLGGVAGLLLFIGLGIFFCVRCRHRRRQAERMSQIKRLLSEKKTCQCPHRFQKTCSPI
                     "
     CDS             2331..3044
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     primer_bind     complement(3082..3099)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3088..3312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3358..3786
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3445..3464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3655..3676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3795..3815)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3800..4129
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3980..4115
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4042..4061
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4196..4990
                     /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(4250..4269)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4860..4879
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5164..5285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5201..5220)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5255..5274
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5334..5350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5334..5350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5347..5369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5358..5374
                     /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(5382..5388,5389..5406,5407..5412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5427..5448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5565..5582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5736..6321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5816..5835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6492..7283,7284..7352))
                     /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     7115..7134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7353..7457)
                     /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 gtttaaactt aagcttagcc gccaccatga accggggagt cccttttagg cacttgcttc
      961 tggtgctgca actggcgctc ctcccagcag ccactcaggg aaagaaagtg gtgctgggca
     1021 aaaaagggga tacagtggaa ctgacctgta cagcttccca gaagaagagc atacaattcc
     1081 actggaaaaa ctccaaccag ataaagattc tgggaaatca gggctccttc ttaactaaag
     1141 gtccatccaa gctgaatgat cgcgctgact caagaagaag cctttgggac caaggaaact
     1201 tccccctgat catcaagaat cttaagatag aagactcaga tacttacatc tgtgaagtgg
     1261 aggaccagaa ggaggaggtg caattgctag tgttcggatt gactgccaac tctgacaccc
     1321 acctgcttca ggggcagagc ctgaccctga ccttggagag cccccctggt agtagcccct
     1381 cagtgcaatg taggagtcca aggggtaaaa acatacaggg ggggaagacc ctctccgtgt
     1441 ctcagctgga gctccaggat agtggcacct ggacatgcac tgtcttgcag aaccagaaga
     1501 aggtggagtt caaaatagac atcgtggtgc tagctttcca gaaggcctcc agcatagtct
     1561 ataagaaaga gggggaacag gtggagttct ccttcccact cgcctttaca gttgaaaagc
     1621 tgacgggcag tggcgagctg tggtggcagg cggagagggc ttcctcctcc aagtcttgga
     1681 tcacctttga cctgaagaac aaggaagtgt ctgtaaaacg ggttacccag gaccctaagc
     1741 tccagatggg caagaagctc ccgctccacc tcaccctgcc ccaggccttg cctcagtatg
     1801 ctggctctgg aaacctcacc ctggcccttg aagcgaaaac aggaaagttg catcaggaag
     1861 tgaacctggt ggtgatgaga gccactcagc tccagaaaaa tttgacctgt gaggtgtggg
     1921 gacccacctc ccctaagctg atgctgagct tgaaactgga gaacaaggag gcaaaggtct
     1981 cgaagcggga gaaggcggtg tgggtgctga accctgaggc ggggatgtgg cagtgtctgc
     2041 tgagtgactc gggacaggtc ctgctggaat ccaacatcaa ggttctgccc acatggtcca
     2101 ccccggtgca gccaatggcc ctgattgtgc tggggggcgt cgccggcctc ctgcttttca
     2161 ttgggctagg catcttcttc tgtgtcaggt gccggcaccg aaggcgccaa gcagagcgga
     2221 tgtctcagat caagagactc ctcagtgaga agaagacctg ccagtgtcct caccggtttc
     2281 agaagacatg tagccccatt gggggggggg ggtccggggg gggggggtcc atggtgtcta
     2341 agggcgaaga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga
     2401 acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga
     2461 ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta
     2521 ccagcttcat gtacggcagc agaaccttca tcaaccacac ccagggcatc cccgacttct
     2581 ttaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg
     2641 gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca
     2701 agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct
     2761 gggaggccaa caccgagatg ctgtaccccg ctgacggcgg cctggaaggc agaaccgaca
     2821 tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttcaagacc acatacagat
     2881 ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac cacagactgg
     2941 aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca
     3001 gatactgcga cctccctagc aaactggggc acaaacttaa ttgactcgag tctagagggc
     3061 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3121 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3181 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3241 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3301 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3361 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3421 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3481 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3541 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3601 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3661 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3721 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3781 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3841 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3901 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3961 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4021 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4081 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4141 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4201 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4261 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4321 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4381 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4441 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4501 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4561 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4621 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4681 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4741 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4801 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4861 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4921 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4981 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5041 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5101 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5161 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5221 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5281 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5341 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5401 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5461 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5521 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5581 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5641 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5701 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5761 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5821 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5881 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5941 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6001 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6061 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6121 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6181 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6241 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     6301 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6361 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6421 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6481 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6541 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6601 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6661 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6721 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6781 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6841 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6901 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6961 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7021 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7081 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7141 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7201 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7261 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7321 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7381 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7441 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
