LOCUS       sequence-420354-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes EV-D68 MO/14/18947 3ABCD optimized for expression in 
            mammalian transcription/translation extracts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  TITLE     Differential Inhibition of Intra- and Inter-molecular Protease 
            Cleavages by Antiviral Compounds
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5867)
  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            "Unpublished"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_3ABCD
FEATURES             Location/Qualifiers
     source          1..5867
                     /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(3215..3241)
                     /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(3215..3241)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3249..3710
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     3637..3654
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3760..3779)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3761..3779)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3786..3803)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3786..3802)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3795..3817)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3943..4398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4030..4049)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4240..4261
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4467..4486)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4586..4608
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4646..4664)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4732..4836
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4837..4905,4906..5697)
                     /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(5055..5074)
                     /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 ggaacgaatc taaccatttc cgtctaagac
      961 tagagtatgc aggtagtgtc aatgctcttc ccacaggcac acttctgatt tttctgcaaa
     1021 attcattata agcctcctct ccattatgcc atgcgaggta gcacaaagac ctaacatgat
     1081 cctgtgtgtt tctgggatct ttagtccatc taatagattc atgtatctct ttcattggca
     1141 tcactggatg tatgagaaag gggtattgat catctgctct gaaatatctt tttaagaaag
     1201 ttacattttc ccaatttgtg tcaataaaac tggttccttt gtctgctggt gtcattacta
     1261 atccatactg tttacctgct tctgccagca aacctggatc aatcttatgt gggtagctag
     1321 caataacatc atccccatag gcaatcattt tgaactggtc caggtctatg cctttgtaaa
     1381 ccctaattaa taaagttctt atgattatat tgttgatcat agtgttgaat atgctggtgc
     1441 ctgaagaacc agagggcatt ccaccattaa ctatgtattt tttgtcttta tataaatgta
     1501 ctgaatgaca caaataatct ataaaagacg tttgatgtgt gtaacctaac ttaattagaa
     1561 ctttctttaa acaggcaaac cacactggtg acaaactagc atcataacca gtgtagtcaa
     1621 aagcaaagat ttctccatct aacaaaatag ggatttttga ccaaaatata tctggatcac
     1681 aaccaactgc actaccagtt gctgtacctg gattgttgtg aaatgtggcg taaaggtttc
     1741 caaaggcaac tctcatagca actgagtcat tcaaggaact ggcctcaatc aggcgtgatt
     1801 tccctttttc aactttttct cttgatctaa gctcatcttt tacaaaggtt acaaaaggta
     1861 ggtcaactcc atatttctct aacatttttg tcatttcact ggtgtctcta gtatgtctat
     1921 taaatatatc ccttttcttc ttcccttgta gtaagtaagg gaatcctgca ctagtagtta
     1981 agtctaatgc ctcaaggcca tccattccat acatggcact ttccaggggt atgggatcca
     2041 cactgatgtc taatggttct aaacacccca cataatgatc cactgcctct tccatgtact
     2101 catccattaa cataattttg ttacctgtgt actttgagaa aatggcctcc tcgaaatctg
     2161 ttttaagcct aggatctttt ggattgagaa ctgctggttc ctttgaacct tcaaaaactt
     2221 gatggaaaac actaggttgg agtttagtct ttgccggtgc gttaatgcac accccactct
     2281 tttcactact aactatctca ccttgtgtat cggaaaagta agagtgtagt agcattgctg
     2341 caaatccttg agctccattt ccacctacat gtattcctat caccttacct gtagttgtca
     2401 ccacaccacc acactggcca gctcttgttg ggaagttata cattaaaatg cggtgcgtcg
     2461 gtgtaccacc taggttcaag aagccataat tggtgacttg tccaactggg atatacatat
     2521 ttgggaattt tgatgtatgt acgctaagca cagcgtcatt gtaatcatcc tcatatctgg
     2581 gcagaaaatg tctgatatct ctaaattttt gattacggtc taatttgact atggttatct
     2641 ctaagtttgt atcagtcaag tctctaagtg cacacgcatc taaaacttta gtctctacat
     2701 cattaatgta aatggtttct ccaacagatg cgtgtgtggg gatgaccgct accctatcat
     2761 atacacccag catggtgaac tcaccctttt cagtccttgc aatgacggta tttttcttca
     2821 ttatggcttg tgcaaaatcg aaccctggtc cttgcacttt agctgttctg agagagggta
     2881 ctttaggttt aggattgggg attcctgtgt atggaccctg aatgccagca aaaagtttgt
     2941 atataacata aactacacca gctattgata caaaggtggc aatggcttgt agagtaataa
     3001 aagctctact aatgtgtttt tctactatta gctcatttga tgggtgtact acaatccatc
     3061 ctttcttttg gcaataatcc ctaacttctt gagaatccac tgacctaaga aggtcattta
     3121 tagcatcagg agctggtgta tctggggtga ctgaaatttt gatctcttta aactgtggag
     3181 gcccgaattc ggatcccata tgggtggtgg ccagggcgta gtcgggcacg tcgtaggggt
     3241 aggccatatt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt cggggggcct
     3301 agacgttttt taacctcgac taaacacatg taaagcatgt gcaccgaggc cccagatcag
     3361 atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat acgcttgagg
     3421 tgagccattt gactctttcc acaactatcc aactcacaac gtggcactgg ggttgtgccg
     3481 cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg tcgccaggtg
     3541 gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa gaagcttcca
     3601 gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg agaggggaaa
     3661 gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc acattgccaa
     3721 aagacggcaa tatggtggaa aataaccgga attaattcgc cctatagtga gtcgtattac
     3781 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3841 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3901 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     3961 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4021 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4081 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4141 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4201 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4261 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4321 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4381 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4441 acaccgcata tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4501 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4561 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4621 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4681 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4741 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4801 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4861 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4921 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4981 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5041 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5101 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5161 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5221 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5281 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5341 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5401 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5461 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5521 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5581 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5641 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5701 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5761 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5821 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
