LOCUS       sequence-443924-        5229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express separately the Rotavirus SA11 NSP3 protein and C1C-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5229)
  TITLE     Rotavirus Nonstructural Protein (NSP) plasmids
REFERENCE   2  (bases 1 to 5229)
  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-NSP3SA11-P2A-C1C-EGFP
FEATURES             Location/Qualifiers
     source          1..5229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..74
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1052..1108
                     /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             join(1379..1381,1382..2095)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1379..1381,1382..2092)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1421..1442)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1682..1701)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2029..2050
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2374..2392)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2388..2435
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2503..2521)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2504..2521)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2811..2830)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3040..3059)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3159..3181
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3219..3237)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3305..3409
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3410..3478,3479..4270)
                     /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(3628..3647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4441..5029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4930..4949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5183..5200
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcaggggg atatcgatcc cgggttaata
       61 cgactcacta taggcattta atgcttttca gtggttgatg ctcaagatgg agtctacgca
      121 acagatggcc gtctcaatta ttaactcttc ttttgaagct gcagttgtag ctgcaacctc
      181 agctcttgag aatatgggaa tagaatatga ttatcaggat atatattcta gagtaaagaa
      241 taaatttgat tttgtgatgg acgattctgg tgttaaaaat aatctgattg gtaaagcaat
      301 aactattgat caagctttga ataataaatt tggatctgct ataagaaata gaaactggct
      361 tgctgatact tctagagcag ctaaattaga tgaggatgta aacaaactaa gaatgatgtt
      421 atcatcaaaa ggaattgatc aaaaaatgag agttttaaac gcatgcttca gtgtaaaaag
      481 aatacctgga aaatcatcat ctattattaa atgcacaaaa ttgatgcgtg ataaattgga
      541 acgtggtgaa gttgaagtgg atgattcatt tgtggatgaa aaaatggaag tggataccat
      601 tgactggaaa tcgcgctatg agcaattgga gcaaaggttt gaatcattga aatccagggt
      661 aaatgaaaaa tataataatt gggtgttgaa agcaagaaaa atgaatgaaa atatgcattc
      721 tcttcaaaat gtcatctctc aacagcaagc acatatagct gagcttcaag tgtacaataa
      781 taaactagaa cgtgatttgc aaaataaaat tggatccctt acttcttcga ttgaatggta
      841 tttaagatca atggaattag accctgaaat aaaggcagac attgaacagc aaattaactc
      901 aattgatgcg ataaatccat tgcacgcttt tgatgactta gaatcagtaa tacgtaattt
      961 gatatctgat tatgacaaat tattccttat gttcaaagga ttaatacaga gatgtaatta
     1021 tcaatattca tttggttgcg aaggaagcgg agcgaccaac tttagcctgc tgaaacaggc
     1081 gggcgatgtg gaagaaaacc cgggcccggc cgacaaggtc ctgaaggaga agagaaagct
     1141 gtttatccgt tccatgggtg aaggtacaat aaatggctta ctggatgaat tattacagac
     1201 aagggtgctg aacaaggaag agatggagaa agtaaaacgt gaaaatgcta cagttatgga
     1261 taagacccga gctttgattg actccgttat tccgaaaggg gcacaggcat gccaaatttg
     1321 catcacatac atttgtgaag aagacagtta cctggcaggg acgctgggac tctcagcagt
     1381 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1441 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1501 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1561 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1621 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1681 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1741 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1801 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1861 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1921 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1981 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2041 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaccatt
     2101 ttgatacatg ttgaacaatc aaatacagtg ttagtatgtt gtcatctatg cataaccctc
     2161 tatgagcaca atagttaaaa gctaacactg tcaaaaacct aaatggctat aggggcgtta
     2221 tgtggccggg tcggcatggc atctccacct cctcgcggtc cgacctgggc atccgaagga
     2281 ggacgtcgtc cactcggatg gctaagggag agctcaaaaa aaaggatccg gctgctaaca
     2341 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     2401 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     2461 cgagatcctc tagagtcgac ctgcaggcat gcaagcttgt attctatagt gtcacctaaa
     2521 tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta acgacaatat
     2581 gtacaagcct aattgtgtag catctggctt actgaagcag accctatcat ctctctcgta
     2641 aactgccgtc agagtcggtt tggttggacg aaccttctga gtttctggta acgccgtccc
     2701 gcacccggaa atggtcagcg aaccaatcag cagggtcatc gctagccaga tcctctacgc
     2761 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     2821 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     2881 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcacca
     2941 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cttaatgcag
     3001 gagtcgcata agggagagcg tcgaatggtg cactctcagt acaatctgct ctgatgccgc
     3061 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3121 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3181 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3241 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3301 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3361 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3421 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3481 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3541 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3601 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3661 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3721 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3781 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3841 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3901 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3961 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4021 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4081 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4141 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4201 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4261 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4321 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4381 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4441 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4501 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4561 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4621 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4681 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4741 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4801 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4861 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4921 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4981 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5041 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5101 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5161 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     5221 acgcaaacc
//
