LOCUS       sequence-218061-        7887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for mammalian expression of HCMV protein UL136. Expresses 
            only the 19kda isoform of UL136. Also expresses EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7887)
  AUTHORS   Caviness K, Cicchini L, Rak M, Umashankar M, Goodrum F
  TITLE     Complex expression of the UL136 gene of human cytomegalovirus 
            results in multiple protein isoforms with unique roles in 
            replication.
  JOURNAL   J Virol. 2014 Dec;88(24):14412-25. doi: 10.1128/JVI.02711-14. Epub 
            2014 Oct 8.
  PUBMED    25297993
REFERENCE   2  (bases 1 to 7887)
  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: 
            "10.1128/JVI.02711-14"; journalName: "J Virol"; date: "2014-12"; 
            volume: "88"; issue: "24"; pages: "14412-25"
COMMENT     Sequence Label: pCIG1_UL136_19kDa_EE
FEATURES             Location/Qualifiers
     source          1..7887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1798..1915
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2005..2208
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2159..2179
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2205..2229
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2630..2647
                     /codon_start=1
                     /product="epitope tag from middle T antigen of mouse 
                     polyoma virus"
                     /label=Glu-Glu tag
                     /translation="EYMPME"
     misc_feature    join(2664..3237,3238..3240,3241..3249)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2831..2848)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3058..3077
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3250..3252,3253..3255,3256..3966)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3250..3252,3253..3255,3256..3966)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3295..3316)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3556..3575)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3903..3924
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3982..4570
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4035..4055)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4453..4464)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4644..4877
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4949..5070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4986..5005)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5040..5059
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5089..5224
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5262..5278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5262..5278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5275..5297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5286..5302
                     /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(5310..5316,5317..5334,5335..5340))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5355..5376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5493..5510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5664..6252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6423..7214,7215..7283))
                     /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     7046..7065
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7284..7388)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7456..7474
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7512..7534)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7634..7653
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7847..7869
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7861..7878
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7862..7878
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc atggatgtgc ccgacccgga actcggcgac ccggcccgcc ggccgttgaa
     2341 cggggccatg tactacggca gcggctgtcg cttcgacacg gtggaaatgg tggacgagac
     2401 gagacccgcg ccgccggcgc tgtcatcgcc cgaaaccggc gacgatagca acgacgacgc
     2461 ggttgccggc ggaggtgctg gcggggtaac atcatccgcg actcgtacga cgtcgtcgaa
     2521 cgcgctgctg ccagaatgga tggatgcggt acatgtggcg gtccaagccg ccgttcaagc
     2581 gaccgtgcaa gtaagtggcc cgcgggagaa cgccgtatct cccgctacgg agtacatgcc
     2641 catggagtaa ggatccaatt ccgcccctct ccctcccccc cccctaacgt tactggccga
     2701 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     2761 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     2821 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     2881 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     2941 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     3001 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     3061 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     3121 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac
     3181 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     3241 gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3301 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     3361 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3421 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3481 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3541 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3601 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3661 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3721 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3781 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3841 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3901 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3961 tacaagtcta gataagtcga caatcaacct ctggattaca aaatttgtga aagattgact
     4021 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4081 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4141 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4201 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4261 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4321 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4381 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4441 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4501 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4561 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     4621 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct
     4681 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     4741 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     4801 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     4861 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     4921 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     4981 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5041 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     5101 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5161 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5221 ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg
     5281 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5341 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5401 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5461 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5521 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5581 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5641 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5701 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5761 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5821 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5881 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5941 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6001 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6061 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6121 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6181 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6241 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6301 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6361 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6421 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6481 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6541 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6601 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6661 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6721 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6781 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6841 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6901 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6961 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7021 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7081 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7141 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7201 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7261 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7321 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7381 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     7441 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     7501 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     7561 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     7621 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     7681 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     7741 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     7801 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7861 tgtaaaacga cggccagtgc caagctg
//
