LOCUS       sequence-420356-        4495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ncodes EV-D68 MO/14/18947 3ABC optimized for expression in mammalian
            transcription/translation extracts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4495)
  TITLE     Differential Inhibition of Intra- and Inter-molecular Protease 
            Cleavages by Antiviral Compounds
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4495)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pT7CFE1_EVD68_3ABC
FEATURES             Location/Qualifiers
     source          1..4495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(789..836)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     832..850
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(908..925)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1843..1869)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             complement(1843..1869)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1877..2338
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     2265..2282
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(2388..2407)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2389..2407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2414..2431)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2414..2430)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2423..2445)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2571..3026
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2658..2677)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2868..2889
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3095..3114)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3214..3236
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3274..3292)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3360..3464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3465..3533,3534..4325)
                     /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     complement(3683..3702)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcccgga tatagttcct
      781 cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
      841 tgctcagcgg actagtgttt aaactttttt tttttttttt tttttttttt ttttcagtca
      901 gatctcagtg gtggtggtgg tggtgctcga gtgtgtatcg gaaaagtaag agtgtagtag
      961 cattgctgca aatccttgag ctccatttcc acctacatgt attcctatca ccttacctgt
     1021 agttgtcacc acaccaccac actggccagc tcttgttggg aagttataca ttaaaatgcg
     1081 gtgcgtcggt gtaccaccta ggttcaagaa gccataattg gtgacttgtc caactgggat
     1141 atacatattt gggaattttg atgtatgtac gctaagcaca gcgtcattgt aatcatcctc
     1201 atatctgggc agaaaatgtc tgatatctct aaatttttga ttacggtcta atttgactat
     1261 ggttatctct aagtttgtat cagtcaagtc tctaagtgca cacgcatcta aaactttagt
     1321 ctctacatca ttaatgtaaa tggtttctcc aacagatgcg tgtgtgggga tgaccgctac
     1381 cctatcatat acacccagca tggtgaactc acccttttca gtccttgcaa tgacggtatt
     1441 tttcttcatt atggcttgtg caaaatcgaa ccctggtcct tgcactttag ctgttctgag
     1501 agagggtact ttaggtttag gattggggat tcctgtgtat ggaccctgaa tgccagcaaa
     1561 aagtttgtat ataacataaa ctacaccagc tattgataca aaggtggcaa tggcttgtag
     1621 agtaataaaa gctctactaa tgtgtttttc tactattagc tcatttgatg ggtgtactac
     1681 aatccatcct ttcttttggc aataatccct aacttcttga gaatccactg acctaagaag
     1741 gtcatttata gcatcaggag ctggtgtatc tggggtgact gaaattttga tctctttaaa
     1801 ctgtggaggc ccgaattcgg atcccatatg ggtggtggcc agggcgtagt cgggcacgtc
     1861 gtaggggtag gccatattat catcgtgttt ttcaaaggaa aaccacgtcc ccgtggttcg
     1921 gggggcctag acgtttttta acctcgacta aacacatgta aagcatgtgc accgaggccc
     1981 cagatcagat cccatacaat ggggtacctt ctgggcatcc ttcagcccct tgttgaatac
     2041 gcttgaggtg agccatttga ctctttccac aactatccaa ctcacaacgt ggcactgggg
     2101 ttgtgccgcc tttgcaggtg tatcttatac acgtggcttt tggccgcaga ggcacctgtc
     2161 gccaggtggg gggttccgct gcctgcaaag ggtcgctaca gacgttgttt gtcttcaaga
     2221 agcttccaga ggaactgctt ccttcacgac attcaacaga ccttgcattc ctttggcgag
     2281 aggggaaaga cccctaggaa tgctcgtcaa gaagacaggg ccaggtttcc gggccctcac
     2341 attgccaaaa gacggcaata tggtggaaaa taaccggaat taattcgccc tatagtgagt
     2401 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2461 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2521 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     2581 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2641 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2701 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2761 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2821 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2881 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2941 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3001 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     3061 ggtatttcac accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3121 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3181 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3241 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     3301 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3361 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3421 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3481 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3541 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3601 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3661 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3721 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3781 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3841 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3901 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3961 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4021 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4081 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4141 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4201 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4261 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4321 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4381 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
