LOCUS       sequence-443935-        5101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Rotavirus SA11 delta103C NSP1 protein fused with the 
            sfGFP (Pedelacq et al., 2006).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5101)
  TITLE     Rotavirus Nonstructural Protein (NSP) plasmids
REFERENCE   2  (bases 1 to 5101)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..5101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(522..540)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(625..642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(796..1384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(876..895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1555..2346,2347..2415))
                     /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     2178..2197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2416..2520)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2588..2606
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2644..2666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2766..2785
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2995..3014
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3304..3322
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3304..3321
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(3390..3437)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3433..3451
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3690..4403)
                     /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     4345..4373
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 atttctcata taacccaaaa acataataaa gattattttc atcaaactta aatgcagcag
       61 catttaatgt aattgggagc agcaaatgat tataccattc aattttatat tgattcctac
      121 attctctttg ctttatcatt ctttcaaatt ttctaattac tttatcatta attggaaata
      181 ttatattgta caagtctatg atacactgta atttgtcttt ggtaattgga tgtttcacag
      241 ttctaagctt aagcaaatga ggttcattgt caaggaagca tcttctgttc tgaacacacc
      301 atttgcaaac atgcggcaat gagcaaccat aacaatgggt taaatcagct atttgacaac
      361 attctaaaca ccacccctta attttcgcat caggaactgg catccaaata gaatttgcac
      421 caatgttgca taatctccga tttaaagcag ttaatctacg ataatgaaag catgcatctt
      481 taaaagtagc catggctaac acaagacttt tcaaaaaaaa gcctatagtg agtcgtatta
      541 acccgggatc gatatccccc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      601 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1441 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1501 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1621 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1741 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1981 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2221 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2401 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2461 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2521 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2581 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2641 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2701 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2761 ctatgcggca tcagagcaga ttgtactgag agtgcaccat tcgacgctct cccttatgcg
     2821 actcctgcat taagaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     2881 ggaatggtgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     2941 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     3001 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     3061 tccggcgtag aggatctggc tagcgatgac cctgctgatt ggttcgctga ccatttccgg
     3121 gtgcgggacg gcgttaccag aaactcagaa ggttcgtcca accaaaccga ctctgacggc
     3181 agtttacgag agagatgata gggtctgctt cagtaagcca gatgctacac aattaggctt
     3241 gtacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
     3301 acgatttagg tgacactata gaatacaagc ttgcatgcct gcaggtcgac tctagaggat
     3361 ctcgatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc
     3421 ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg
     3481 gctttgttag cagccggatc cttttttttg agctctccct tagccatccg agtggacgac
     3541 gtcctccttc ggatgcccag gtcggaccgc gaggaggtgg agatgccatg ccgacccggt
     3601 tcacagtatt ttgccagcta ggcgctactc tagtctggac ggtcttgtga tggctaaata
     3661 ctgtataatt agatagtgac ataatttcat ttgtagagtt catccatgcc atgtgtaatc
     3721 ccagcagcag ttacatcctc aagaaggacc atgtggtcac gcttttcgtt gggatctttc
     3781 gaaagaacag attgtgtcga gaggtaatgg ttgtctggta aaaggacagg gccatcgcca
     3841 attggagtat tttgttgata atggtctgct agttgaacgg aaccatcttc aacgttgtgg
     3901 cgaattttga agtacgcttt gattccattc ttttgtttgt ctgccgtgat gtatacattg
     3961 tgtgagttaa agttgtactc cagtttgtgt ccgagaatgt ttccatcttc tttaaaatca
     4021 atacctttta actcgatacg attaacaagg gtatcacctt caaacttgac ttcagcacgc
     4081 gtcttgtagg tcccgtcatc tttgaaagat atagtgcgtt cctgtacata accttcgggc
     4141 atggcactct tgaaaaagtc atgccgtttc atgtgatccg gataacggga aaagcataga
     4201 acaccatagg tcagagtagt gacaagtgtt ggccacggaa caggtagttt tccagtagtg
     4261 caaataaatt taagggtgag ttttccgttt gtagcatcac cttcaccctc tccacggaca
     4321 gaaaatttgt gcccattaac atcaccatct aattcaacaa gaattgggac aactccagtg
     4381 aaaagttctt ctcctttgct catgtgatta aacacttctt caattgtttg atccatgtga
     4441 ttagtaaaca aactccaaat taatctttta attaaaggat atatgttttc aacagaacta
     4501 caatgtccaa cgttaacatt acttttataa caatccttaa taaattccat caataaatta
     4561 tacatatcac gaatccgcca atcaatttta ctaagaaaat attttatatt acaccattta
     4621 cagtactgga tatcggaagc attgcctggg tgattagatg ccacgtagtt aggctgacaa
     4681 tgatgtccca actcaaatat tgtcattttc aaccttccaa aattattatc atgagcatag
     4741 aaccgagtat acttactcgt catcattgca gcaattaatt tattcttatc attttttaaa
     4801 tcagaataaa gctccaattc atctctccaa ttaaattcag ttacatttct gtatattgat
     4861 cttcctggcg tttttgtata agtattaaaa atcacagttt ctttagtaaa atctgaaaag
     4921 taaatttgct ttagttttga tgatggcagt cttgataatt ggaagtattt ctttgcgtaa
     4981 tggtttctca actctattgg tagattggac attcttgtaa agttaatatg atcaagcaat
     5041 aatttatcaa attcatttat aaagttaaca attctatatg gagcatatat atcactgact
     5101 g
//
