LOCUS       sequence-443936-        3945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Rotavirus SA11 NSP4 fused with the PEST degradation 
            sequence from mouse ornithine decarboxylase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3945)
  TITLE     Rotavirus Nonstructural Protein (NSP) plasmids
REFERENCE   2  (bases 1 to 3945)
  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"
COMMENT     Sequence Label: pT7-NSP4SA11-hPEST
FEATURES             Location/Qualifiers
     source          1..3945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     108..127
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     337..356
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        646..664
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     646..663
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(732..779)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     775..793
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1125..1244)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     promoter        complement(1809..1827)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1912..1929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2083..2671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2163..2182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2842..3633,3634..3702))
                     /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     3465..3484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3703..3807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3875..3893
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(3931..3945,1..8))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
       61 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
      121 gattgtactg agagtgcacc attcgacgct ctcccttatg cgactcctgc attaagaagc
      181 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcaaggagat
      241 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
      301 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
      361 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg
      421 gctagcgatg accctgctga ttggttcgct gaccatttcc gggtgcggga cggcgttacc
      481 agaaactcag aaggttcgtc caaccaaacc gactctgacg gcagtttacg agagagatga
      541 tagggtctgc ttcagtaagc cagatgctac acaattaggc ttgtacatat tgtcgttaga
      601 acgcggctac aattaataca taaccttatg tatcatacac atacgattta ggtgacacta
      661 tagaatacaa gcttgcatgc ctgcaggtcg actctagagg atctcgatcc ggatatagtt
      721 cctcctttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt
      781 tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga
      841 tccttttttt tgagctctcc cttagccatc cgagtggacg acgtcctcct tcggatgccc
      901 aggtcggacc gcgaggaggt ggagatgcca tgccgacccg gtcacattaa gaccgttcct
      961 tccattaacg tccggtgctg accggattgg ttaaacggga ttaaggcttg agccacacgc
     1021 gatatggctg actgtggctt ctcagtcagc catcaggtcc gatggtgctt actgtaaaga
     1081 actccgccgc ttccgaggac agtcgacggc agctcaacct cttagacgtt gatgcgagct
     1141 gaagcacaag cggctgggtg ccggtccata ccgctttctt gtgcgcagga cataggcaat
     1201 gtaccggcgg cttgctcttc aacctcaggc ggaaagccgt gagacattgc tgcagtcact
     1261 tctcttggtt cataaggatt ttttccactt tcccattctt ctagcgttct cacgtttttt
     1321 tgattgatct cttttgtcat atctatttca cctgtcgttt gcaccgtcaa tttatcgtaa
     1381 atgcgtttaa gcaactctac ttgttcaatt tcacgtgtag tcaatttgtc aatcatttct
     1441 agctggcgtc tcatttcttt gactactctg tccatttgct tttctatctc atctttagta
     1501 gttatctgct ctttataacc tgccaatttt aacaacgtat taaaaattgt tacaatacaa
     1561 tatttcacca ctttatatga acattttgac gttttcaatg caattttcat tgttggaatg
     1621 gatgctttat gtagcgcaaa caaaactgtt aagacagatg ctatataagg aaaatacgcc
     1681 attcctggat cctcaagtat tgtgtgcaat gtattgttca ttagagtgat tacactcaat
     1741 gtataattga ggtcggtaag cttttccatc tttccgcacg cgctctctcg gaacagaact
     1801 tttaaaagcc tatagtgagt cgtattaacc cgggatcgat atccccctgc attaatgaat
     1861 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3841 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3901 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgca
//
