LOCUS       sequence-403314-        6092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for human CD4 coding sequence tagged with the mOrange FP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6092)
  AUTHORS   Iliopoulou M, Nolan R, Alvarez L, Watanabe Y, Coomer CA, 
            Jakobsdottir GM, Bowden TA, Padilla-Parra S
  TITLE     A dynamic three-step mechanism drives the HIV-1 pre-fusion reaction.
  JOURNAL   Nat Struct Mol Biol. 2018 Sep;25(9):814-822. doi: 
            10.1038/s41594-018-0113-x. Epub 2018 Aug 27.
  PUBMED    30150645
REFERENCE   2  (bases 1 to 6092)
  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"; doi: 
            "10.1038/s41594-018-0113-x"; journalName: "Nat Struct Mol Biol"; 
            date: "2018-09"; volume: "25"; issue: "9"; pages: "814-822"
COMMENT     Sequence Label: hCD4-mOrange
FEATURES             Location/Qualifiers
     source          1..6092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        180..483
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        484..687
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     638..658
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(738..812,813..2111)
                     /codon_start=1
                     /product="human CD4 cell surface glycoprotein"
                     /label=CD4
                     /translation="MNRGVPFRHLLLVLQLALLPAATQGKKVVLGKKGDTVELTCTASQ
                     KKSIQFHWKNSNQIKILGNQGSFLTKGPSKLNDRADSRRSLWDQGNFPLIIKNLKIEDS
                     DTYICEVEDQKEEVQLLVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRGKNI
                     QGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFQKASSIVYKKEGEQVEFS
                     FPLAFTVEKLTGSGELWWQAERASSSKSWITFDLKNKEVSVKRVTQDPKLQMGKKLPLH
                     LTLPQALPQYAGSGNLTLALEAKTGKLHQEVNLVVMRATQLQKNLTCEVWGPTSPKLML
                     SLKLENKEAKVSKREKAVWVLNPEAGMWQCLLSDSGQVLLESNIKVLPTWSTPVQPMAL
                     IVLGGVAGLLLFIGLGIFFCVRCRHRRRQAERMSQIKRLLSEKKTCQCPHRFQKTCSPI
                     "
     regulatory      2160..2169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2166..2876
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2579..2598
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2999..3120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3036..3055)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3090..3109
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3127..3582)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3264..3285)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3476..3495
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3609..3713
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3715..4072
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3715..3906
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3738..3758)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3923..4058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4107..4901
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4107..4901
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4161..4180)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4771..4790
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5089..5108)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5133..5180
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5509..6092,1..5)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5998..6017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
       61 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
      121 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      181 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      241 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      301 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      361 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      421 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      481 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      541 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      601 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      661 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      721 cggactcaga tctcgagatg aaccggggag tcccttttag gcacttgctt ctggtgctgc
      781 aactggcgct cctcccagca gccactcagg gaaagaaagt ggtgctgggc aaaaaagggg
      841 atacagtgga actgacctgt acagcttccc agaagaagag catacaattc cactggaaaa
      901 actccaacca gataaagatt ctgggaaatc agggctcctt cttaactaaa ggtccatcca
      961 agctgaatga tcgcgctgac tcaagaagaa gcctttggga ccaaggaaac ttccccctga
     1021 tcatcaagaa tcttaagata gaagactcag atacttacat ctgtgaagtg gaggaccaga
     1081 aggaggaggt gcaattgcta gtgttcggat tgactgccaa ctctgacacc cacctgcttc
     1141 aggggcagag cctgaccctg accttggaga gcccccctgg tagtagcccc tcagtgcaat
     1201 gtaggagtcc aaggggtaaa aacatacagg gggggaagac cctctccgtg tctcagctgg
     1261 agctccagga tagtggcacc tggacatgca ctgtcttgca gaaccagaag aaggtggagt
     1321 tcaaaataga catcgtggtg ctagctttcc agaaggcctc cagcatagtc tataagaaag
     1381 agggggaaca ggtggagttc tccttcccac tcgcctttac agttgaaaag ctgacgggca
     1441 gtggcgagct gtggtggcag gcggagaggg cttcctcctc caagtcttgg atcacctttg
     1501 acctgaagaa caaggaagtg tctgtaaaac gggttaccca ggaccctaag ctccagatgg
     1561 gcaagaagct cccgctccac ctcaccctgc cccaggcctt gcctcagtat gctggctctg
     1621 gaaacctcac cctggccctt gaagcgaaaa caggaaagtt gcatcaggaa gtgaacctgg
     1681 tggtgatgag agccactcag ctccagaaaa atttgacctg tgaggtgtgg ggacccacct
     1741 cccctaagct gatgctgagc ttgaaactgg agaacaagga ggcaaaggtc tcgaagcggg
     1801 agaaggcggt gtgggtgctg aaccctgagg cggggatgtg gcagtgtctg ctgagtgact
     1861 cgggacaggt cctgctggaa tccaacatca aggttctgcc cacatggtcc accccggtgc
     1921 agccaatggc cctgattgtg ctggggggcg tcgccggcct cctgcttttc attgggctag
     1981 gcatcttctt ctgtgtcagg tgccggcacc gaaggcgcca agcagagcgg atgtctcaga
     2041 tcaagagact cctcagtgag aagaagacct gccagtgccc tcaccggttt cagaagacat
     2101 gtagccccat tttaaagctt cgaattctgc agtcgacggt accgcgggcc cgggatcccg
     2161 ccaccatggt gagcaagggc gaggagaata acatggccat catcaaggag ttcatgcgct
     2221 tcaaggtgcg catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     2281 agggccgccc ctacgagggc tttcagaccg ctaagctgaa ggtgaccaag ggtggccccc
     2341 tgcccttcgc ctgggacatc ctgtcccctc agttcaccta cggctccaag gcctacgtga
     2401 agcaccccgc cgacatcccc gactacttca agctgtcctt ccccgagggc ttcaagtggg
     2461 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     2521 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     2581 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     2641 acggcgccct gaagggcgag atcaagatga ggctgaagct gaaggacggc ggccactaca
     2701 cctccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     2761 tcgtcggcat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2821 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaaagcg
     2881 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2941 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3001 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3061 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3121 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3181 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3241 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3301 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3361 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3421 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3481 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3541 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3721 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3841 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3901 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3961 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4021 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4081 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4141 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4201 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4261 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4321 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4381 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4441 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4501 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4561 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4621 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4681 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4741 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4801 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4861 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4921 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4981 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5041 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5101 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5161 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5221 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5281 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5341 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5401 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5461 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5641 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6061 tttgtgatgc tcgtcagggg ggcggagcct at
//
