LOCUS       sequence-443928-        5167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express separately the Rotavirus SA11 delta103C NSP1 protein and the
            sfGFP (Pedelacq et al., 2006).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5167)
  TITLE     Rotavirus Nonstructural Protein (NSP) plasmids
REFERENCE   2  (bases 1 to 5167)
  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-NSP1delta103CSA11-P2A-GFP
FEATURES             Location/Qualifiers
     source          1..5167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        51..155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(156..224,225..1016)
                     /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(374..393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1187..1775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1676..1695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1929..1946
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2031..2049
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3278..3334
                     /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             3335..4048
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVLCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     YFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     DVTAAGITHGMDELYK"
     primer_bind     complement(3365..3393)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(4287..4305)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4301..4348
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(4416..4434)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4417..4434)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4724..4743)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4953..4972)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5072..5094
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5132..5150)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
       61 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      121 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      181 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      241 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      301 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      361 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      421 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      481 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      541 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
      601 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
      661 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
      721 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      781 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      841 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
      901 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
      961 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1021 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1081 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1141 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1201 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1261 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1321 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1381 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1441 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1501 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1561 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1621 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1681 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1741 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1801 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1861 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1921 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     1981 tccccgcgcg ttggccgatt cattaatgca gggggatatc gatcccgggt taatacgact
     2041 cactataggc ttttttttga aaagtcttgt gttagccatg gctactttta aagatgcatg
     2101 ctttcattat cgtagattaa ctgctttaaa tcggagatta tgcaacattg gtgcaaattc
     2161 tatttggatg ccagttcctg atgcgaaaat taaggggtgg tgtttagaat gttgtcaaat
     2221 agctgattta acccattgtt atggttgctc attgccgcat gtttgcaaat ggtgtgttca
     2281 gaacagaaga tgcttccttg acaatgaacc tcatttgctt aagcttagaa ctgtgaaaca
     2341 tccaattacc aaagacaaat tacagtgtat catagacttg tacaatataa tatttccaat
     2401 taatgataaa gtaattagaa aatttgaaag aatgataaag caaagagaat gtaggaatca
     2461 atataaaatt gaatggtata atcatttgct gctcccaatt acattaaatg ctgctgcatt
     2521 taagtttgat gaaaataatc tttattatgt ttttgggtta tatgagaaat cagtcagtga
     2581 tatatatgct ccatatagaa ttgttaactt tataaatgaa tttgataaat tattgcttga
     2641 tcatattaac tttacaagaa tgtccaatct accaatagag ttgagaaacc attacgcaaa
     2701 gaaatacttc caattatcaa gactgccatc atcaaaacta aagcaaattt acttttcaga
     2761 ttttactaaa gaaactgtga tttttaatac ttatacaaaa acgccaggaa gatcaatata
     2821 cagaaatgta actgaattta attggagaga tgaattggag ctttattctg atttaaaaaa
     2881 tgataagaat aaattaattg ctgcaatgat gacgagtaag tatactcggt tctatgctca
     2941 tgataataat tttggaaggt tgaaaatgac aatatttgag ttgggacatc attgtcagcc
     3001 taactacgtg gcatctaatc acccaggcaa tgcttccgat atccagtact gtaaatggtg
     3061 taatataaaa tattttctta gtaaaattga ttggcggatt cgtgatatgt ataatttatt
     3121 gatggaattt attaaggatt gttataaaag taatgttaac gttggacatt gtagttctgt
     3181 tgaaaacata tatcctttaa ttaaaagatt aatttggagt ttgtttacta atcacatgga
     3241 tcaaacaatt gaagaagtgt ttaatcacgg aagcggagct actaacttca gcctgctgaa
     3301 gcaggctgga gacgtggagg agaaccctgg acctatgagc aaaggagaag aacttttcac
     3361 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
     3421 ccgtggagag ggtgaaggtg atgctacaaa cggaaaactc acccttaaat ttatttgcac
     3481 tactggaaaa ctacctgttc cgtggccaac acttgtcact actctgacct atggtgttct
     3541 atgcttttcc cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc
     3601 cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggacct acaagacgcg
     3661 tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa aaggtattga
     3721 ttttaaagaa gatggaaaca ttctcggaca caaactggag tacaacttta actcacacaa
     3781 tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gcgtacttca aaattcgcca
     3841 caacgttgaa gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg
     3901 cgatggccct gtccttttac cagacaacca ttacctctcg acacaatctg ttctttcgaa
     3961 agatcccaac gaaaagcgtg accacatggt ccttcttgag gatgtaactg ctgctgggat
     4021 tacacatggc atggatgaac tctacaaatg aaattatgtc actatctaat tatacagtat
     4081 ttagccatca caagaccgtc cagactagag tagcgcctag ctggcaaaat actgtgaacc
     4141 gggtcggcat ggcatctcca cctcctcgcg gtccgacctg ggcatccgaa ggaggacgtc
     4201 gtccactcgg atggctaagg gagagctcaa aaaaaaggat ccggctgcta acaaagcccg
     4261 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     4321 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatcgagatc
     4381 ctctagagtc gacctgcagg catgcaagct tgtattctat agtgtcacct aaatcgtatg
     4441 tgtatgatac ataaggttat gtattaattg tagccgcgtt ctaacgacaa tatgtacaag
     4501 cctaattgtg tagcatctgg cttactgaag cagaccctat catctctctc gtaaactgcc
     4561 gtcagagtcg gtttggttgg acgaaccttc tgagtttctg gtaacgccgt cccgcacccg
     4621 gaaatggtca gcgaaccaat cagcagggtc atcgctagcc agatcctcta cgccggacgc
     4681 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     4741 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     4801 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca ccattccttg
     4861 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcttaatg caggagtcgc
     4921 ataagggaga gcgtcgaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4981 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5041 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5101 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5161 aatgtca
//
