LOCUS       sequence-444017-        4382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express separately the short Rotavirus SA11 NSP1 protein (35 amino 
            acids) and the full-length recoded NSP5 protein fused with GS-linker
            and smURFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4382)
  TITLE     Rotavirus Nonstructural Protein (NSP) plasmids
REFERENCE   2  (bases 1 to 4382)
  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: pT7-35aaNSP1SA11-P2A-NSP5(io)-GS-smURFP
FEATURES             Location/Qualifiers
     source          1..4382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     85..107
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(145..163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        231..335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(336..404,405..1196)
                     /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(554..573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1367..1955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1856..1875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2109..2126
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2211..2230
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2211..2229
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2364..2420
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3045..3446
                     /codon_start=1
                     /product="dimeric small ultra-red fluorescent protein 
                     derived from a cyanobacterial phycobiliprotein (Rodriguez 
                     et al., 2016)"
                     /label=smURFP
                     /translation="MAKTSEQRVNIATLLTENKKKIVDKASQDLWRRHPDLIAPGGIAF
                     SQRDRALCLRDYGWFLHLITFCLLAGDKGPIESIGLISIREMYNSLGVPVPAMMESIRC
                     LKEASLSLLDEEDANETAPYFDYIIKAMS"
     primer_bind     complement(3682..3700)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3696..3743
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(3811..3829)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3812..3829)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4119..4138)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4348..4367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
       61 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      121 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
      181 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      541 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      721 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
      781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
      841 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
      901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1081 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1201 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1261 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1441 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1501 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1561 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1621 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1681 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1741 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1801 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1861 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1921 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1981 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2041 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2101 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2161 tccccgcgcg ttggccgatt cattaatgca gggggatatc gatcccgggt taatacgact
     2221 cactataggg cttttttttg aaaagtcttg tgttagccat ggctactttt aaagatgcat
     2281 gctttcatta tcgtagatta actgctttaa atcggagatt atgcaacatt ggtgcaaatt
     2341 ctatttggat gccagttcct gatgctacta acttcagcct gctgaagcag gctggagacg
     2401 tggaggagaa ccctggacct atgagtttaa gtatagatgt tacatcactt ccctcgatcc
     2461 cctcaacaat atataaaaat gaaagttcaa gtacaacaag tacactatcc ggaaaatcta
     2521 taggaagaag tgaacaatat ataagcccag atgcagaagc ttttaataaa tatatgctaa
     2581 gtaaatcacc tgaagatata gggccaagtg atagtgcatc aaacgatccc ctcacaagtt
     2641 tttcaatacg aagtaacgca gtaaaaacta atgccgatgc aggtgtaagc atggactcct
     2701 cagcacaatc aagaccttca tctaacgtag gatgtgatca agtcgatttc tcattaaaca
     2761 aaggattaaa agttaaagca aatttagatt catctataag tataagcaca gatacaaaaa
     2821 aagaaaaaag taaacaaaat cataaaagtc gaaaacacta tccaagaata gaagccgaaa
     2881 gtgatagcga tgattatgta ctagacgaca gcgattcaga tgatggaaaa tgtaaaaatt
     2941 gcaaatacaa aaaaaaatat tttgcactaa gaatgagaat gaaacaagtt gccatgcaat
     3001 taattgaaga tttaggttct ggatccggtt ctggaagtgg atccatggct aagacttccg
     3061 aacagagggt gaacattgct acactgctga cagaaaataa gaagaaaatc gtggataagg
     3121 cttcccagga tctgtggcgg agacacccag acctgatcgc accaggagga attgctttct
     3181 ctcagaggga ccgcgctctg tgcctgcgag attacggctg gttcctgcat ctgatcacct
     3241 tttgtctgct ggccggagat aagggcccca tcgagtctat tgggctgatc agtattcgag
     3301 aaatgtataa ctcactggga gtgcccgtcc ctgcaatgat ggagagcatt agatgcctga
     3361 aagaagccag cctgtccctg ctggacgaag aggacgccaa cgagaccgca ccctactttg
     3421 attacattat taaggctatg agctaaaatt atgtcactat ctaattatac agtatttagc
     3481 catcacaaga ccgtccagac tagagtagcg cctagctggc aaaatactgt gaaccgggtc
     3541 ggcatggcat ctccacctcc tcgcggtccg acctgggcat ccgaaggagg acgtcgtcca
     3601 ctcggatggc taagggagag ctcaaaaaaa aggatccggc tgctaacaaa gcccgaaagg
     3661 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     3721 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatcg agatcctcta
     3781 gagtcgacct gcaggcatgc aagcttgtat tctatagtgt cacctaaatc gtatgtgtat
     3841 gatacataag gttatgtatt aattgtagcc gcgttctaac gacaatatgt acaagcctaa
     3901 ttgtgtagca tctggcttac tgaagcagac cctatcatct ctctcgtaaa ctgccgtcag
     3961 agtcggtttg gttggacgaa ccttctgagt ttctggtaac gccgtcccgc acccggaaat
     4021 ggtcagcgaa ccaatcagca gggtcatcgc tagccagatc ctctacgccg gacgcatcgt
     4081 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     4141 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     4201 ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcaccatt ccttgcggcg
     4261 gcggtgctca acggcctcaa cctactactg ggctgcttct taatgcagga gtcgcataag
     4321 ggagagcgtc gaatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4381 gc
//
