LOCUS       sequence-263256-        6229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for HCMV glycoprotein L (TB40/E strain)
            with FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6229)
  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 6229)
  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-HCMVgL-FLAG (TB40/E)
FEATURES             Location/Qualifiers
     source          1..6229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             862..885
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(929..946)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    935..1159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1205..1633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1292..1311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1502..1523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1642..1662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1647..1976
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1827..1962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1889..1908
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2043..2837
                     /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(2097..2116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2707..2726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3011..3132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3048..3067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3102..3121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3181..3197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3181..3197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3194..3216)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3205..3221
                     /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(3229..3235,3236..3253,3254..3259))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3274..3295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3412..3429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3583..4168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3663..3682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4339..5130,5131..5199))
                     /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     4962..4981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5200..5304)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5379..5398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5570..5949
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5950..6153
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6104..6124
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6198..6217
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6198..6216
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gctagcgcca ccatgtgcag aaggccggat tgcggtttta gcttttcccc aggaccagta
       61 gtgctcctgt ggtgctgtct gctccttcca atagtatcta gtgtggctgt cagcgtagcg
      121 ccaactgccg ctgagaaggt ccccgctgag tgtcctgagt tgacccggcg ctgccttctc
      181 ggcgaagtct tccagggaga caaatatgag agctggttgc gccctttggt gaatgtaacc
      241 ggcagaaacg gtcccctgag tcaattgatt cggtacaggc cagtaacgcc tgaggccgca
      301 aattctgtgc ttctggacga tgctttcttg gacaccctgg cactgctgta taacaacccg
      361 gatcagctcc gggcgcttct cacgcttttg agcagcgaca cagccccacg atggatgacc
      421 gtcatgcgcg gctactccga atgtggggat ggaagtccgg ctgtatacac atgtgtggac
      481 gatctgtgtc gcgggtacga tcttacaagg ctgtcctatg gccgctccat attcacagag
      541 cacgttctcg gcttcgagct ggtgcctccg tctctgttca atgtcgtcgt agcgattcga
      601 aatgaggcca cacggaccaa ccgcgcagtt cggctgccag ttagcaccgc ggctgctcct
      661 gaggggatta ctcttttcta tggtctttac aacgcggtta aggaattctg tcttcggcac
      721 caactcgacc cacccctctt gcgccatctg gacaagtact acgcaggttt gcctcctgag
      781 ctgaaacaaa caagggttaa cctgccagcg cacagtcgat acgggccaca ggcggtcgat
      841 gcaaggagcg ggggatccgg agattataaa gatgacgatg acaagtgata actcgagtct
      901 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
      961 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1021 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1081 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     1141 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     1201 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1261 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1321 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1381 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1441 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1501 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1561 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1621 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1681 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     1741 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1801 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1861 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     1921 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     1981 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     2041 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2101 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2161 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2221 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2281 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2341 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2401 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2461 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2521 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     2581 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2641 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2701 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2761 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     2821 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     2881 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     2941 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3001 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3061 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3121 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     3181 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3241 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3301 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3361 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3421 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4021 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     4141 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4201 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4261 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4321 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4381 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4441 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4501 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4561 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4621 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4681 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4741 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4801 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4861 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4921 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4981 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5041 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5101 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5161 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5221 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5281 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     5341 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     5401 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     5461 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     5521 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     5581 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5641 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     5701 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     5761 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     5821 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     5881 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     5941 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     6001 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     6061 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     6121 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     6181 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctg
//
