LOCUS       sequence-258941-        3804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRDV5 plasmid containing a loop Sybody for the generation of a qPCR 
            standard curve.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3804)
  AUTHORS   Zimmermann I, Egloff P, Hutter CAJ, Kuhn BT, Brauer P, Newstead S, 
            Dawson RJP, Geertsma ER, Seeger MA
  TITLE     Generation of synthetic nanobodies against delicate proteins.
  JOURNAL   Nat Protoc. 2020 Apr 8. pii: 10.1038/s41596-020-0304-x. doi: 
            10.1038/s41596-020-0304-x.
  PUBMED    32269381
REFERENCE   2  (bases 1 to 3804)
  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"; journalName: "Nat Protoc.
            2020 Apr 8. pii: 10.1038/s41596-020-0304-x. doi: 
            10.1038/s41596-020-0304-x."
COMMENT     Sequence Label: pRDV5 containing Sb-loop
FEATURES             Location/Qualifiers
     source          1..3804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             69..91
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1442..1460)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1456..1503
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1539..1994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1626..1645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1836..1857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2021..2125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2126..2194,2195..2986)
                     /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(2344..2363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3157..3744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3645..3664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acccgacacc atcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gaaataattt tgtttaactt taagaaggag atatatccat gggtagtcag gttcagctgg
      121 ttgagagcgg tggtggcctg gtccaagctg gcggttcgct gcgtctgagc tgcgccgcaa
      181 gcggtttccc ggtgagcagc agcacgatga cctggtatcg tcaggcaccg ggcaaagaac
      241 gtgagtgggt cgcggcgatt tccagctctg gtagcaccac gacctacgca gattctgtta
      301 agggccgctt taccatcagc cgcgacaacg cgaagaatac ggtctatttg cagatgaata
      361 gcctgaaacc ggaagatacc gcggtttact actgtaacgt gaaagacagc ggtagctcca
      421 gcagctccta cgactattgg ggccaaggta cccaagtgac tgtgagcgca aagctttata
      481 tggcctcggg ggccgaattc ggatctggtg gccagaagca agctgaagag gcggcagcga
      541 aagcggcggc agatgctaaa gcgaaggccg aagcagatgc taaagctgcg gaagaagcag
      601 cgaagaaagc ggctgcagac gcaaagaaaa aagcagaagc agaagccgcc aaagccgcag
      661 ccgaagcgca gaaaaaagcc gaggcagccg ctgcggcact gaagaagaaa gcggaagcgg
      721 cagaagcagc tgcagctgaa gcaagaaaga aagcggcaac tgaagctgct gaaaaagcca
      781 aagcagaagc tgagaagaaa gcggctgctg aaaaggctgc agctgataag aaagcggcag
      841 cagagaaagc tgcagccgac aaaaaagcag cagaaaaagc ggctgctgaa aaggcagcag
      901 ctgataagaa agcagcggca gaaaaagccg ccgcagacaa aaaagcggca gcggcaaaag
      961 ctgcagctga aaaagccgct gcagcaaaag cggccgcaga ggcagatgat attttcggtg
     1021 agctaagctc tggtaagaat gcaccgaaaa cggggggagg ggcgaaaggg aacaatgctt
     1081 cgcctgccgg gagtggtaat actaaaaaca atggcgcatc aggggccgat atcaataact
     1141 atgccgggca gattaaatct gctatcgaaa gtaagttcta tgacgcatcg tcctatgcag
     1201 gcaaaacctg tacgctgcgc ataaaactgg cacccgatgg tatgttactg gatatcaaac
     1261 ctgaaggtgg cgatcccgca ctttgtcagg ctgcgttggc agcagctaaa cttgcgaaga
     1321 taccgaaacc accaagccag gcagtatatg aagtgttcaa aaacgcgcca ttggacttca
     1381 aaccgtagta gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1441 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1501 ttgctgaaag gaggaactat atccggatcg agatccccac gcgccctgta gcggcgcatt
     1561 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1621 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1681 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1741 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1801 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1861 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1921 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1981 aacgcttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2041 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2101 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2161 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2221 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2281 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2341 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2401 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2461 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2521 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2581 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2641 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2701 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2761 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2821 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2881 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2941 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3001 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3061 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3121 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3181 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3241 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3301 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3361 atacctcgct ctgctaatcc tgttaccagt ggctctgcca gtggcgataa gtcgtgtctt
     3421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3781 ttttgctggc cttttgctca catg
//
