LOCUS       sequence-263263-        6751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for HCMV glycoprotein O (TB40/E strain)
            with an 8his tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6751)
  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 6751)
  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-8his (TB40/E)
FEATURES             Location/Qualifiers
     source          1..6751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1111..1122)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1384..1407
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(1451..1468)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1457..1681
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1727..2155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1814..1833)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2024..2045
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2164..2184)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2169..2498
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2349..2484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2411..2430
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2565..3359
                     /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(2619..2638)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3229..3248
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3533..3654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3570..3589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3624..3643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3703..3719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3703..3719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3716..3738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3727..3743
                     /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(3751..3757,3758..3775,3776..3781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3796..3817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3934..3951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4105..4690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4185..4204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4861..5652,5653..5721))
                     /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     5484..5503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5722..5826)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5901..5920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6092..6471
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6472..6675
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6626..6646
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6720..6739
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6720..6738
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gctagcgcca ccatgaagtg ggtgactttc atcagtcttt tgttcttgtt tagctcagct
       61 tacagcaagg ttgtgaggcc tccaggaaga tattggcttg gaaccgtcct gtccacaatc
      121 ggaaaacaaa aacttgacaa atttaaattg gagatactga agcaacttga gcgcgagcca
      181 tataccaaat atttcaatat gactaggcaa cacgtcaaaa accttactat gaatatgacg
      241 cagtttccgc aatattacat ccttgcagga cccattagaa atgactccat aacctatctt
      301 tggttcgatt tttacagtac ccaactgcga aagccagcga agtacgtcta cagccaatac
      361 aatcacacgg cgaaaacgat cactttccgc ccacctagct gtgggaccgt accgtctatg
      421 acctgcttga gtgagatgtt gaatgtttca aagcgaaacg ataccggtga gcaaggatgc
      481 gggaacttta caacctttaa tcccatgttc ttcaacgttc ccagatggaa taccaaactc
      541 tacgtcggcc ctactaaagt caatgtagat agtcagacta tctacttttt gggattgaca
      601 gcattgctct tgaggtatgc tcagcggaat tgcacacata gtttttacct cgtgaatgct
      661 atgagtagga atctgttccg agtcccgaag tatattaacg gaactaaact gaagaacacg
      721 atgagaaagc tgaaacgcaa gcaagccccg gtgaaagagc agcttgaaaa gaaaacaaag
      781 aaaagtcaga gcactactac gccttacttc tcctatacga cgagcacagc actgaatgtt
      841 actacaaacg ccacctacag agtcactacg tccgcaaagc gaatccctac gagcactatc
      901 gcgtacaggc cggattccag tttcatgaag agcattatgg cgactcagtt gagagatttg
      961 gcaacatggg tctacactac gcttcgctat cggaatgagc cattttgtaa acctgatcga
     1021 aaccggacag ctgttagtga attcatgaaa aatacgcacg tgttgattag aaatgaaacg
     1081 ccttatacta tttatggaac gcttgacatg tcttccctct attacaacga gactatgtcc
     1141 gtagaaaatg agacggcgtc tgacaacaac gagaccactc cgacttcccc gagtactagg
     1201 tttcaaaaaa ccttcattga ccctttgtgg gactatctcg acagcttgct ctttctcgat
     1261 aagattcgca atttttcttt gcaactcccg gcctacggta acctcacgcc gcctgaacat
     1321 cggagagcag ttaatctttc aaccttgaat agtctctggt ggtggctcca gggtggatcc
     1381 ggtcaccatc accaccacca tcatcattga taactcgagt ctagagggcc cgtttaaacc
     1441 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1501 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1561 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1621 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1681 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     1741 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1801 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1861 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1921 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1981 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2041 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2101 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2161 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2221 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2281 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2341 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2401 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2461 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     2521 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2581 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2641 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     2701 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     2761 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     2821 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     2881 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     2941 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3001 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3061 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3121 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3181 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3241 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3301 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3361 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3421 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3481 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     3541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3661 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     3721 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3781 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3841 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3901 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3961 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4021 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4081 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4141 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4201 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4261 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4321 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4381 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4441 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4501 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4561 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4621 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4681 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4741 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4801 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4861 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4921 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4981 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5041 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5101 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5161 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5221 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5281 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5341 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5401 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5461 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5521 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5581 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5641 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5701 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5761 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5821 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     5881 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     5941 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     6001 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     6061 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     6121 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     6181 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     6241 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     6301 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     6361 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     6421 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     6481 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     6541 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     6601 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     6661 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     6721 aatacgactc actataggga gacccaagct g
//
