LOCUS       sequence-271690-        7459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for HCMV glycoprotein O (TB40/E strain)
            fused to superfolder GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7459)
  AUTHORS   Park J, Gill KS, Aghajani AA, Heredia JD, Choi H, Oberstein A, 
            Procko E
  TITLE     Engineered receptors for human cytomegalovirus that are orthogonal 
            to normal human biology.
  JOURNAL   PLoS Pathog. 2020 Jun 19;16(6):e1008647. doi: 
            10.1371/journal.ppat.1008647. eCollection 2020 Jun.
  PUBMED    32559251
REFERENCE   2  (bases 1 to 7459)
  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: "PLoS 
            Pathog."; date: "2020-06-19"; volume: "16(6):e1008647. doi"; pages: 
            " 10.1371/journal.ppat.1008647. eCollection 2020 Jun"
COMMENT     Sequence Label: pcDNA3-HCMVgO-sfGFP (TB40/E)
FEATURES             Location/Qualifiers
     source          1..7459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1771..1782)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2062..2775
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             2062..2772
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2089..2117)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2712..2732
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2816..2833)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2822..3046
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3092..3520
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3179..3198)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3389..3410
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3529..3549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3534..3863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3714..3849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3776..3795
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3930..4724
                     /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(3984..4003)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4594..4613
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4898..5019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4935..4954)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4989..5008
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5068..5084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5068..5084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5081..5103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5092..5108
                     /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(5116..5122,5123..5140,5141..5146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5161..5182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5299..5316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5470..6058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5550..5569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6229..7020,7021..7089))
                     /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     6852..6871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7090..7194)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7269..7288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgcca ccatgaagtg ggtgactttc atcagtcttt tgttcttgtt tagctcagct
      721 tacagcaagg ttgtgaggcc tccaggaaga tattggcttg gaaccgtcct gtccacaatc
      781 ggaaaacaaa aacttgacaa atttaaattg gagatactga agcaacttga gcgcgagcca
      841 tataccaaat atttcaatat gactaggcaa cacgtcaaaa accttactat gaatatgacg
      901 cagtttccgc aatattacat ccttgcagga cccattagaa atgactccat aacctatctt
      961 tggttcgatt tttacagtac ccaactgcga aagccagcga agtacgtcta cagccaatac
     1021 aatcacacgg cgaaaacgat cactttccgc ccacctagct gtgggaccgt accgtctatg
     1081 acctgcttga gtgagatgtt gaatgtttca aagcgaaacg ataccggtga gcaaggatgc
     1141 gggaacttta caacctttaa tcccatgttc ttcaacgttc ccagatggaa taccaaactc
     1201 tacgtcggcc ctactaaagt caatgtagat agtcagacta tctacttttt gggattgaca
     1261 gcattgctct tgaggtatgc tcagcggaat tgcacacata gtttttacct cgtgaatgct
     1321 atgagtagga atctgttccg agtcccgaag tatattaacg gaactaaact gaagaacacg
     1381 atgagaaagc tgaaacgcaa gcaagccccg gtgaaagagc agcttgaaaa gaaaacaaag
     1441 aaaagtcaga gcactactac gccttacttc tcctatacga cgagcacagc actgaatgtt
     1501 actacaaacg ccacctacag agtcactacg tccgcaaagc gaatccctac gagcactatc
     1561 gcgtacaggc cggattccag tttcatgaag agcattatgg cgactcagtt gagagatttg
     1621 gcaacatggg tctacactac gcttcgctat cggaatgagc cattttgtaa acctgatcga
     1681 aaccggacag ctgttagtga attcatgaaa aatacgcacg tgttgattag aaatgaaacg
     1741 ccttatacta tttatggaac gcttgacatg tcttccctct attacaacga gactatgtcc
     1801 gtagaaaatg agacggcgtc tgacaacaac gagaccactc cgacttcccc gagtactagg
     1861 tttcaaaaaa ccttcattga ccctttgtgg gactatctcg acagcttgct ctttctcgat
     1921 aagattcgca atttttcttt gcaactcccg gcctacggta acctcacgcc gcctgaacat
     1981 cggagagcag ttaatctttc aaccttgaat agtctctggt ggtggctcca gggtggatcc
     2041 ggtgggtccg gtagtggtgg ttccaaagga gaagaacttt tcactggagt tgtcccaatt
     2101 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa
     2161 ggtgatgcta caaacggaaa actcaccctt aaatttattt gcactactgg aaaactacct
     2221 gttccatggc caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat
     2281 ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     2341 gaacgcacta tatctttcaa agatgacggg acctacaaga cgcgtgctga agtcaagttt
     2401 gaaggtgata cccttgttaa tcgtatcgag ttaaaaggta ttgattttaa agaagatgga
     2461 aacattctcg gacacaaact ggagtacaac tttaactcac acaatgtata catcacggca
     2521 gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacgt tgaagatggt
     2581 tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     2641 ttaccagaca accattacct gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag
     2701 cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat
     2761 gagctctaca aatgataact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     2821 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2881 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2941 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3001 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3061 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3121 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3181 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3241 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3301 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3361 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3421 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3481 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3541 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3601 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3661 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3721 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3781 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3841 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3901 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3961 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4021 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4081 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4141 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4201 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4261 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4321 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4381 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4441 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4501 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4561 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4621 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4681 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4741 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4801 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4861 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4921 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4981 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5041 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5101 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5161 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5221 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5281 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6301 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6361 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6421 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6481 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6541 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6601 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6661 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6721 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6781 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6841 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6901 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6961 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7021 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7081 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7201 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     7261 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     7321 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     7381 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     7441 cgggccagat atacgcgtt
//
