LOCUS       sequence-218062-        7980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for mammalian expression of HCMV protein UL136. Expresses 
            only the 23kda isoform of UL136. Also expresses EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7980)
  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 7980)
  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_23kDa_Myc
FEATURES             Location/Qualifiers
     source          1..7980
                     /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             2714..2740
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(2757..3330,3331..3333,3334..3342)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2924..2941)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3151..3170
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3343..3345,3346..3348,3349..4059)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3343..3345,3346..3348,3349..4059)
                     /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(3388..3409)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3649..3668)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3996..4017
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4075..4663
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4128..4148)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4546..4557)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4737..4970
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5042..5163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5079..5098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5133..5152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5182..5317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5355..5371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5355..5371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5368..5390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5379..5395
                     /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(5403..5409,5410..5427,5428..5433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5448..5469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5586..5603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5757..6345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6516..7307,7308..7376))
                     /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     7139..7158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7377..7481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7549..7567
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7605..7627)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7727..7746
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7940..7962
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7954..7971
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7955..7971
                     /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 atgctccacg atctattttg cggctgtcat tatcctgaga agtgccgtcg
     2341 gcaccacgag cggcagagaa gcagacggcg agccgcggat gtgcccgacc cggaactcgg
     2401 cgacccggcc cgccggccgt tgaacggggc catgtactac ggcagcggct gtcgcttcga
     2461 cacggtggaa atggtggacg agacgagacc cgcgccgccg gcgctgtcat cgcccgaaac
     2521 cggcgacgat agcaacgacg acgcggttgc cggcggaggt gctggcgggg taacatcatc
     2581 cgcgactcgt acgacgtcgt cgaacgcgct gctgccagaa tggatggatg cggtacatgt
     2641 ggcggtccaa gccgccgttc aagcgaccgt gcaagtaagt ggcccgcggg agaacgccgt
     2701 atctcccgct acgtacccat acgatgttcc agattacgcg taaggatcca attccgcccc
     2761 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     2821 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     2881 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     2941 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     3001 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     3061 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     3121 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     3181 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     3241 tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg
     3301 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag
     3361 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3421 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3481 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3541 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3601 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3661 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3721 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3781 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     3841 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3901 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3961 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4021 gccgccggga tcactctcgg catggacgag ctgtacaagt ctagataagt cgacaatcaa
     4081 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4141 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4201 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4261 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4321 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4381 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4441 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4501 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4561 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4621 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc
     4681 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     4741 agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg ggtctctctg
     4801 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     4861 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     4921 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     4981 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     5041 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5101 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5161 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg
     5221 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5281 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agacggccca
     5341 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5401 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5461 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5521 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6181 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7501 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7561 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     7621 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     7681 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     7741 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     7801 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7861 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     7921 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
