LOCUS       sequence-263252-        7543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for HCMV glycoprotein H (TB40/E strain)
            extracellular domain with 8his purification tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7543)
  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 7543)
  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-HCMVgH-8his (TB40/E)
FEATURES             Location/Qualifiers
     source          1..7543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2176..2199
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2243..2260)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2249..2473
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2519..2947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2606..2625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2816..2837
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2956..2976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2961..3290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3141..3276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3203..3222
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3357..4151
                     /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(3411..3430)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4021..4040
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4325..4446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4362..4381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4416..4435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4495..4511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4495..4511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4508..4530)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4519..4535
                     /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(4543..4549,4550..4567,4568..4573))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4588..4609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4726..4743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4897..5482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4977..4996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5653..6444,6445..6513))
                     /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     6276..6295
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6514..6618)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6693..6712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6884..7263
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7264..7467
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7418..7438
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7512..7531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7512..7530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gctagcgcca ccatgcgacc agggcttccg ttttatttga ccgtatttgc tgtttatctg
       61 ctttcccact tgcctagtca acgctatggg gccgatgctg cgtctgaggc ccttgaccct
      121 cacgcattcc acctcctgct taacacatac ggccggccca ttcgctttct ccgagagaat
      181 acgacccaat gcacttacaa ctcatctctc cgcaacagca cagtagtccg cgagaacgcc
      241 atatctttca acttctttca atcatacaat cagtactatg ttttccacat gccccggtgc
      301 cttttcgcgg gcccattggc tgaacaattc cttaatcaag tagatctcac ggagacgctt
      361 gaacggtatc aacagcggct caacacgtat gcgttggtaa gcaaggattt ggcctcatat
      421 cggagcttta gtcagcagtt gaaggcgcag gacagcctcg ggcaacagcc aacaacggtc
      481 cccccgccga tcgacctcag cattcctcac gtttggatgc cccctcaaac tactcctcat
      541 gattggaaag gttcccatac cacatccggc ctgcacaggc ctcattttaa ccaaacatgt
      601 atcctgtttg atggacatga cctcctgttc tcaacggtaa cgccatgtct ccatcaaggt
      661 ttctatctga tggatgagct caggtacgta aagataacct tgacggaaga ttttttcgtt
      721 gtgactgtgt ccatcgacga cgatacgcca atgctcctga tttttggaca cctcccgaga
      781 gtcctgttta aggcgcccta tcagcgagac aacttcattc tgcgccaaac tgaaaagcat
      841 gaattgctcg tacttgtgaa aaaaacgcaa ctcaaccgac acagctactt gaaggatagc
      901 gattttttgg atgctgcatt ggattttaat tatctggacc tctccgcatt gctgaggaat
      961 tctttccata ggtatgcggt tgacgtgttg aagtctggac gatgtcagat gttggacaga
     1021 aggacggttg agatggcttt cgcatacgct ctcgctcttt ttgcagcagc tagacaggaa
     1081 gaggccggta cagaaattag tatccctaga gcactggata ggcaggcagc tctcttgcag
     1141 attcaagagt ttatgataac ttgtctctct caaacgcctc cacgaaccac gctcctcctc
     1201 tacccgacag cggtagactt ggctaagcga gctctgtgga cgcccgacca aatcacagac
     1261 ataacaagcc ttgttcgctt ggtgtacatc cttagtaagc agaatcaaca acatcttata
     1321 ccccagtggg ctctgcgcca gattgcagat ttcgcgctgc agttgcacaa aacacacctg
     1381 gcatctttcc tgagcgcttt tgctcgacag gagctctacc tcatggggag cttggtacat
     1441 tccatgttgg ttcatacgac ggaacggcga gaaattttca tagtcgaaac cggcctgtgt
     1501 tcacttgcag agctcagcca ttttacacag ctgctggcac acccgcacca tgagtacctc
     1561 tctgaccttt acacaccctg ttctagctct ggaagacggg atcactcctt ggagaggctt
     1621 accaggctct tcccggatgc gaccgtaccg gcaacggtac cggctgctct ttctatcctc
     1681 agcacaatgc aaccaagtac tttggagact tttccggatc tcttctgcct tccactcgga
     1741 gagagtttta gcgcgttgac agtatctgaa cacgtgagct acgttgtaac taaccaatat
     1801 ctcatcaaag gcatcagtta tcctgtttcc acaacagtgg ttggccagtc actcataata
     1861 actcaaaccg actcccaatc aaaatgtgag ctcacacgaa acatgcacac gacacatagc
     1921 ataacggccg ccctcaatat atctcttgaa aactgcgctt tttgccaaag tgcgcttttg
     1981 gagtatgatg acacgcaggg agtaattaac atcatgtaca tgcacgattc agacgatgtc
     2041 ttgttcgcgc tggaccccta taacgaggtt gtggtgtcaa gcccgcgaac ccattacctc
     2101 atgctgctta aaaacggtac ggttttggag gtaacggacg tcgtggtaga cgctacagat
     2161 agcggcggat ccggtcacca tcaccaccac catcatcatt gataactcga gtctagaggg
     2221 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2281 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2341 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2401 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2461 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2521 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2581 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2641 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2701 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2761 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2821 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2881 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2941 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3301 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3361 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3421 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3481 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3541 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3601 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3661 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3721 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3781 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3841 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3901 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3961 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4021 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4081 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4141 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4201 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4261 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4321 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4441 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4681 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5401 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     5461 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5521 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5581 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5641 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5701 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5761 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5821 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5881 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5941 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6001 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6061 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6121 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6181 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6241 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6301 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6361 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6421 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6481 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6541 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6601 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     6661 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6721 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     6781 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     6841 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     6901 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     6961 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7021 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7081 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7141 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7201 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7261 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     7321 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     7381 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     7441 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
     7501 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctg
//
