LOCUS       sequence-443927-        4974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express separately the Rotavirus SA11 NSP3 protein and the original 
            enhanced GFP (Yang et al., 1996) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4974)
  TITLE     Rotavirus Nonstructural Protein (NSP) plasmids
REFERENCE   2  (bases 1 to 4974)
  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-F2A-EGFP
FEATURES             Location/Qualifiers
     source          1..4974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..168
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        253..271
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1246..1311
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             join(1312..1314,1315..1317,1318..2028)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1312..1314,1315..1317,1318..2028)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1357..1378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1618..1637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1965..1986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2316..2334)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2330..2377
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2445..2463)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2446..2463)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2753..2772)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2982..3001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3101..3123
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3161..3179)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3247..3351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3352..3420,3421..4212)
                     /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(3570..3589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4383..4971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4872..4891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
       61 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      121 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      181 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagggggata
      241 tcgatcccgg gttaatacga ctcactatag gcatttaatg cttttcagtg gttgatgctc
      301 aagatggagt ctacgcaaca gatggccgtc tcaattatta actcttcttt tgaagctgca
      361 gttgtagctg caacctcagc tcttgagaat atgggaatag aatatgatta tcaggatata
      421 tattctagag taaagaataa atttgatttt gtgatggacg attctggtgt taaaaataat
      481 ctgattggta aagcaataac tattgatcaa gctttgaata ataaatttgg atctgctata
      541 agaaatagaa actggcttgc tgatacttct agagcagcta aattagatga ggatgtaaac
      601 aaactaagaa tgatgttatc atcaaaagga attgatcaaa aaatgagagt tttaaacgca
      661 tgcttcagtg taaaaagaat acctggaaaa tcatcatcta ttattaaatg cacaaaattg
      721 atgcgtgata aattggaacg tggtgaagtt gaagtggatg attcatttgt ggatgaaaaa
      781 atggaagtgg ataccattga ctggaaatcg cgctatgagc aattggagca aaggtttgaa
      841 tcattgaaat ccagggtaaa tgaaaaatat aataattggg tgttgaaagc aagaaaaatg
      901 aatgaaaata tgcattctct tcaaaatgtc atctctcaac agcaagcaca tatagctgag
      961 cttcaagtgt acaataataa actagaacgt gatttgcaaa ataaaattgg atcccttact
     1021 tcttcgattg aatggtattt aagatcaatg gaattagacc ctgaaataaa ggcagacatt
     1081 gaacagcaaa ttaactcaat tgatgcgata aatccattgc acgcttttga tgacttagaa
     1141 tcagtaatac gtaatttgat atctgattat gacaaattat tccttatgtt caaaggatta
     1201 atacagagat gtaattatca atattcattt ggttgcgaag ccccagtcaa acagactctc
     1261 aactttgatc tactcaagct ggcaggtgat gtggaatcta accccgggcc catggtgagc
     1321 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1381 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1441 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1501 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1561 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1621 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1681 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1741 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1801 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1861 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1921 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1981 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagga attctaacca
     2041 ttttgataca tgttgaacaa tcaaatacag tgttagtatg ttgtcatcta tgcataaccc
     2101 tctatgagca caatagttaa aagctaacac tgtcaaaaac ctaaatggct ataggggcgt
     2161 tatgtggccg ggtcggcatg gcatctccac ctcctcgcgg tccgacctgg gcatccgaag
     2221 gaggacgtcg tccactcgga tggctaaggg agagctcaaa aaaaaggatc cggctgctaa
     2281 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     2341 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     2401 atcgagatcc tctagagtcg acctgcaggc atgcaagctt gtattctata gtgtcaccta
     2461 aatcgtatgt gtatgataca taaggttatg tattaattgt agccgcgttc taacgacaat
     2521 atgtacaagc ctaattgtgt agcatctggc ttactgaagc agaccctatc atctctctcg
     2581 taaactgccg tcagagtcgg tttggttgga cgaaccttct gagtttctgg taacgccgtc
     2641 ccgcacccgg aaatggtcag cgaaccaatc agcagggtca tcgctagcca gatcctctac
     2701 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     2761 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     2821 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcac
     2881 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcttaatgc
     2941 aggagtcgca taagggagag cgtcgaatgg tgcactctca gtacaatctg ctctgatgcc
     3001 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3061 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3121 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3181 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3241 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3301 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3361 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3421 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3481 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3541 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3601 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3661 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3721 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3781 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3841 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3901 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3961 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4021 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4081 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4141 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4201 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4501 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaac
//
