LOCUS       sequence-420355-        4481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Poliovirus type I Mahoney 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 4481)
  TITLE     Differential Inhibition of Intra- and Inter-molecular Protease 
            Cleavages by Antiviral Compounds
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4481)
  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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..4481
                     /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(1829..1855)
                     /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(1829..1855)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1863..2324
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     2251..2268
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(2374..2393)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2375..2393)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2400..2417)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2400..2416)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2409..2431)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2557..3012
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2644..2663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2854..2875
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3081..3100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3200..3222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3260..3278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3346..3450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3451..3519,3520..4311)
                     /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(3669..3688)
                     /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 gactctgagt gaagtatgat cgcttcaggg
      961 ccgctgcaaa cccgtgtgaa ccgttcccac caacatgcat cccgatgact ttcccagtac
     1021 atgtgatgac tccaccacac tgtcctgctc tggttggaaa gttgtacatt agagtacgag
     1081 cagtttggcg cccaccgaga tttagatatc cctgttcagt cacagcaccg acaggaacat
     1141 acatattggg gtacttgcta gtgttcacga tcaagactcc atcatttgtc tcagtgattt
     1201 gagtaggtat atgtggtcta atgtctctga acttttcatt tctctttaga gtgattatag
     1261 tgatttcaag attggttcct gcttgatctt cgagcgcttt ggcatccaag atctccactt
     1321 ctttgccatc gatcacaatg ctttcaccag gtgaagcgtg ggttggtaaa atagccacgt
     1381 tgtcgtggac tcctaacata gtgaactctc ccttgctagt agttgctgta acaatgtttc
     1441 ttttagccat agccactgcg taatcgaacc ctggtccttg tacctttgct gtccgaatgg
     1501 tgggcacgtt gggttttttg tttggtaaac cagtgtatgc tccctggtgt ccagcaaaca
     1561 gtttatacat gacatagaca actccagcca ctgcggcgaa ggttgtcacc gcttgtagaa
     1621 ttgtcattgc cctgttgatg ttcctttctg tttgaacctg gctggtgatg ttgactatcc
     1681 aacccttctt ctcacagtaa tctctcacct cctgggagtc aactgcttgg agcaagtcat
     1741 tgatacattc aggaggggga ctcgtcttga tgtcaatttt caagtcttta tactggagtg
     1801 gtccggatcc catatgggtg gtggccaggg cgtagtcggg cacgtcgtag gggtaggcca
     1861 tattatcatc gtgtttttca aaggaaaacc acgtccccgt ggttcggggg gcctagacgt
     1921 tttttaacct cgactaaaca catgtaaagc atgtgcaccg aggccccaga tcagatccca
     1981 tacaatgggg taccttctgg gcatccttca gccccttgtt gaatacgctt gaggtgagcc
     2041 atttgactct ttccacaact atccaactca caacgtggca ctggggttgt gccgcctttg
     2101 caggtgtatc ttatacacgt ggcttttggc cgcagaggca cctgtcgcca ggtggggggt
     2161 tccgctgcct gcaaagggtc gctacagacg ttgtttgtct tcaagaagct tccagaggaa
     2221 ctgcttcctt cacgacattc aacagacctt gcattccttt ggcgagaggg gaaagacccc
     2281 taggaatgct cgtcaagaag acagggccag gtttccgggc cctcacattg ccaaaagacg
     2341 gcaatatggt ggaaaataac cggaattaat tcgccctata gtgagtcgta ttacaattca
     2401 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2461 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2521 ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
     2581 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2641 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2701 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2761 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2821 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2881 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2941 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3001 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3061 catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3121 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3181 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3241 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3301 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3361 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3421 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3481 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3541 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3601 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3661 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3721 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3781 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3841 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3901 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3961 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4021 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4081 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4141 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4201 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4261 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4321 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4381 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
