LOCUS       sequence-407379-        6828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Kunjin virus PrME protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  AUTHORS   Strebinger D, Frangieh CJ, Friedrich MJ, Faure G, Macrae RK, Zhang F
  TITLE     Cell type-specific delivery by modular envelope design.
  JOURNAL   Nat Commun. 2023 Aug 23;14(1):5141. doi: 10.1038/s41467-023-40788-8.
  PUBMED    37612276
REFERENCE   2  (bases 1 to 6828)
  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: "Nat 
            Commun."; date: "2023-08-23"; volume: "14(1):5141. doi"; pages: " 
            10.1038/s41467-023-40788-8"
COMMENT     Sequence Label: pHCMV_KUNV_PrME
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          792..1364
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      1419..1428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3533..3552)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3598..3653
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3652..3671)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4011..4027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4011..4027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4024..4046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4035..4051
                     /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(4059..4065,4066..4083,4084..4089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4104..4125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4242..4259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4413..5001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4493..4512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5172..5963,5964..6032))
                     /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     5795..5814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6033..6137)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6163..6618)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6300..6321)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6512..6531
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6745..6767
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6759..6776
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6760..6776
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6786..6805
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6786..6804
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca
       61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaca cgccatccac
      721 gctgttttga cctccataga tgacaccggg accgatccag cctccggtcg accgatcctg
      781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      961 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
     1021 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
     1081 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1141 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1201 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt
     1261 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1321 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc
     1381 tggttgttgt gctgtctcat cattttggca aagaattcgc caccatgggc ggcggcaaga
     1441 ccggaatcgc cttcatgatc ggcctcattg ccggcgtggg cgccgtgaca ctgtccaact
     1501 tccaaggcaa ggtcatgatg acagtgaacg ccaccgacgt caccgacatt atcaccattc
     1561 ctcccgccgc cggcaaaaat ctgtgcatcg tgagagctat ggatgtcgga cacatgtgtg
     1621 acgacaccat cacctatgag tgccccgtgc tgagcgccgg aaatgacccc gaggatatcg
     1681 actgctggtg taccaagctc gccgtctacg tgagatacgg aagatgcaca aagacaagac
     1741 atagcagaag gtctagaagg tctctgacag tgcagacaca tggcgagagc acactgagca
     1801 ataagaaggg cgcttggatg gactccacaa aagccacaag atacctcgtg aaaaccgaga
     1861 gctggattct gagaaatccc ggatacgctc tcgtcgctgc tgtgatcgga tggatgctcg
     1921 gctccaacac aatgcagaga gtggtgtttg ctgtgctgct gctcctcgtc gcccccgcct
     1981 attccttcaa ctgtctgggc atgtccaata gagactttct ggaaggagtg tccggagcca
     2041 catgggtgga tctcgtgctc gagggcgaca gctgcgtcac aatcatgtcc aaggacaaac
     2101 ccaccatcga cgtgaagatg atgaatatgg aggccgccaa cctcgctgag gtgagatcct
     2161 actgttatct cgccaccgtg tccgaactct ccacaaaggc cgcttgcccc accatgggag
     2221 aagcccacaa tgacaaaaga gccgacccca gcttcgtgtg caagcaaggc gtggtggata
     2281 gaggatgggg caatggctgt ggcctcttcg gcaagggatc catcgataca tgcgccaagt
     2341 ttgcttgtag caccaaggcc accggaagaa caatcctcaa ggagaacatc aagtacgagg
     2401 tggccatttt tgtccatgga cccacaaccg tggagagcca cggcaactat ttcacccaga
     2461 ccggcgctgc tcaagctgga agattcagca tcacacccgc cgcccccagc tacacactga
     2521 aactgggaga gtatggcgaa gtgacagtgg actgtgagcc tagatccggc atcgacacct
     2581 ccgcttatta tgtgatgacc gtgggcacca agacattcct cgtgcataga gaatggttca
     2641 tggatctgaa tctgccttgg agctccgccg aaagcaacgt gtggagaaat agggagacac
     2701 tgatggagtt cgaggagccc catgctacca agcagagcgt gatcgctctg ggcagccaag
     2761 agggagctct gcaccaagct ctggccggcg ccatccccgt ggagtttagc tccaacacag
     2821 tgaagctcac atccggccat ctcaaatgta gagtcaagat ggagaaactg cagctgaaag
     2881 gcaccaccta cggagtgtgt agcaaggcct ttaggttcct cggcacaccc gccgataccg
     2941 gccacggaac agtggtcctc gaactgcagt acaccggcac cgatggacct tgcaagattc
     3001 ctatttccag cgtggcctct ctgaatgatc tgacccccgt gggcagactc gtcacagtca
     3061 acccctttgt cagcgtcagc acagccaacg ctaaggtgct gattgaactc gagccccctt
     3121 ttggcgatag ctacatcgtg gtgggaaggg gcgagcaaca gatcaaccat cactggcaca
     3181 agtccggcag cagcatcggc aaagccttca ccgccacact caagggcgcc caaagactgg
     3241 ctgccctcgg agacaccgct tgggatttcg gaagcgtcgg aggcgtgttt acaagcgtcg
     3301 gaaaggctgt ccatcaagtg ttcggcggcg cttttagatc cctcttcgga ggaatgagct
     3361 ggattaccca aggactgctg ggcgctctgc tgctgtggat gggcatcaat gctagagata
     3421 ggtccattgc tctgaccttt ctggccgtgg gaggagtgct gctgtttctg tccgtcaacg
     3481 tgcacgcttg atgtcttcct ggagctccag tcattgacta taacgtctca gatctttttc
     3541 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     3601 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     3661 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     3721 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     3781 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3841 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3901 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3961 ctcttctctt atggagatcc ctcgacggat cggccgcaat tcgtaatcat gtcatagctg
     4021 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc accagattta
     5341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     6181 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     6241 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     6301 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     6361 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     6421 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     6481 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     6541 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     6601 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6661 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6721 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     6781 gcgcgtaata cgactcacta tagggcgaat tggagctcca ccgcggtg
//
