LOCUS       sequence-422575-        4480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Poliovirus type I Mahoney 3ABC with active site mutation 
            optimized for expression in mammalian transcription/translation 
            extracts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4480)
  TITLE     Differential Inhibition of Intra- and Inter-molecular Protease 
            Cleavages by Antiviral Compounds
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4480)
  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            "Unpublished"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_polio_3ABC_C147R
FEATURES             Location/Qualifiers
     source          1..4480
                     /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(907..924)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1828..1854)
                     /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(1828..1854)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1862..2323
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     2250..2267
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(2373..2392)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2374..2392)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2399..2416)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2399..2415)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2408..2430)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2556..3011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2643..2662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2853..2874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3080..3099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3199..3221
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3259..3277)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3345..3449
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3450..3518,3519..4310)
                     /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(3668..3687)
                     /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 tttcagtcag
      901 atctcagtgg tggtggtggt ggtgctcgag actctgagtg aagtatgatc gcttcagggc
      961 cgctgcaaac ccgtgtgaac cgttcccacc aacatgcatc ccgatgactt tcccagtaca
     1021 tgtgatgact ccacctctct gtcctgctct ggttggaaag ttgtacatta gagtacgagc
     1081 agtttggcgc ccaccgagat ttagatatcc ctgttcagtc acagcaccga caggaacata
     1141 catattgggg tacttgctag tgttcacgat caagactcca tcatttgtct cagtgatttg
     1201 agtaggtata tgtggtctaa tgtctctgaa cttttcattt ctctttagag tgattatagt
     1261 gatttcaaga ttggttcctg cttgatcttc gagcgctttg gcatccaaga tctccacttc
     1321 tttgccatcg atcacaatgc tttcaccagg tgaagcgtgg gttggtaaaa tagccacgtt
     1381 gtcgtggact cctaacatag tgaactctcc cttgctagta gttgctgtaa caatgtttct
     1441 tttagccata gccactgcgt aatcgaaccc tggtccttgt acctttgctg tccgaatggt
     1501 gggcacgttg ggttttttgt ttggtaaacc agtgtatgct ccctggtgtc cagcaaacag
     1561 tttatacatg acatagacaa ctccagccac tgcggcgaag gttgtcaccg cttgtagaat
     1621 tgtcattgcc ctgttgatgt tcctttctgt ttgaacctgg ctggtgatgt tgactatcca
     1681 acccttcttc tcacagtaat ctctcacctc ctgggagtca actgcttgga gcaagtcatt
     1741 gatacattca ggagggggac tcgtcttgat gtcaattttc aagtctttat actggagtgg
     1801 tccggatccc atatgggtgg tggccagggc gtagtcgggc acgtcgtagg ggtaggccat
     1861 attatcatcg tgtttttcaa aggaaaacca cgtccccgtg gttcgggggg cctagacgtt
     1921 ttttaacctc gactaaacac atgtaaagca tgtgcaccga ggccccagat cagatcccat
     1981 acaatggggt accttctggg catccttcag ccccttgttg aatacgcttg aggtgagcca
     2041 tttgactctt tccacaacta tccaactcac aacgtggcac tggggttgtg ccgcctttgc
     2101 aggtgtatct tatacacgtg gcttttggcc gcagaggcac ctgtcgccag gtggggggtt
     2161 ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt caagaagctt ccagaggaac
     2221 tgcttccttc acgacattca acagaccttg cattcctttg gcgagagggg aaagacccct
     2281 aggaatgctc gtcaagaaga cagggccagg tttccgggcc ctcacattgc caaaagacgg
     2341 caatatggtg gaaaataacc ggaattaatt cgccctatag tgagtcgtat tacaattcac
     2401 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2461 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     2521 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
     2581 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2641 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2701 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2761 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2821 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2881 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2941 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3001 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3061 atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3121 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3181 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3241 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3301 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3361 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3421 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3481 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3541 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3601 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3661 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3721 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3781 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3841 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3901 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3961 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4021 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4081 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4141 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4201 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4261 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4321 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4381 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4441 actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
