LOCUS       sequence-396563-        4340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nanobody.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4340)
  AUTHORS   Cornish K, Huo J, Jones L, Sharma P, Thrush JW, Abdelkarim S, Kipar 
            A, Ramadurai S, Weckener M, Mikolajek H, Liu S, Buckle I, Bentley E,
            Kirby A, Han X, Laidlaw SM, Hill M, Eyssen L, Norman C, Le Bas A, 
            Clarke J, James W, Stewart JP, Carroll M, Naismith JH, Owens RJ
  TITLE     Structural and functional characterization of nanobodies that 
            neutralize Omicron variants of SARS-CoV-2.
  JOURNAL   Open Biol. 2024 Jun;14(6):230252. doi: 10.1098/rsob.230252. Epub 
            2024 Jun 4.
  PUBMED    38835241
REFERENCE   2  (bases 1 to 4340)
  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: "Open 
            Biol."; date: "2024-06"; volume: "14(6):230252. doi"; pages: " 
            10.1098/rsob.230252. Epub 2024 Jun 4"
COMMENT     Sequence Label: pOPINVHH_H6_His
FEATURES             Location/Qualifiers
     source          1..4340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(126..194,195..986)
                     /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(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1910..1931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1946..1951,1952..1969,1970..1976)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1984..2000
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     sig_peptide     2025..2090
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             2472..2489
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2512..2541
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      complement(3906..4334)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4016..4037)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4228..4247
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc gcttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     1861 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     1921 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     1981 gaattgtgag cggataacaa tttgaattca aggagacagt cataatgaaa tacctattgc
     2041 ctacggcggc cgctggattg ttattactcg cggcccagcc ggccatggcc caggtgcagc
     2101 tggtcgagtc tgggggaggc ttggtgcagc ctggggggtc tctgacactc tcctgtgtag
     2161 cgtcggaatc atctttggcg ccttatcgcg tagcctggtt ccgtcaggcc ccagggaagg
     2221 agcgcgaggg ggtctcatgt attagtcgtg atgcacatcc tactagtaca tactatacag
     2281 cctccgtgaa gggccgattc accatgtcca gagacaatgc caagaacacg gtgtatctgc
     2341 aaatgaacag cctgaaacca tcggacacgg ccgtttatta ctgtgcgaca gatttgggag
     2401 gatactgttc ggattctaat tatccccgcg cctggtgggg ccaggggacc caggtcaccg
     2461 tctcctcagg ccaccatcac catcaccatt aataaccacc atcatggcgc agaacaaaaa
     2521 ctcatctcag aagaggatct gtcttaggcc gaaactgttg aaagttgttt agcaaaacct
     2581 catacagaaa attcatttac taacgtctgg aaagacgaca aaactttaga tcgttacgct
     2641 aactatgagg gctgtctgtg gaatgctaca ggcgttgtgg tttgtactgg tgacgaaact
     2701 cagtgttacg gtacatgggt tcctattggg cttgctatcc ctgaaaatga gggtggtggc
     2761 tctgagggtg gcggttctga gggtggcggt tctgagggtg gcggtactaa acctcctgag
     2821 tacggtgata cacctattcc gggctatact tatatcaacc ctctcgacgg cacttatccg
     2881 cctggtactg agcaaaaccc cgctaatcct aatccttctc ttgaggagtc tcagcctctt
     2941 aatactttca tgtttcagaa taataggttc cgaaataggc agggtgcatt aactgtttat
     3001 acgggcactg ttactcaagg cactgacccc gttaaaactt attaccagta cactcctgta
     3061 tcatcaaaag ccatgtatga cgcttactgg aacggtaaat tcagagactg cgctttccat
     3121 tctggcttta atgaggatcc attcgtttgt gaatatcaag gccaatcgtc tgacctgcct
     3181 caacctcctg tcaatgctgg cggcggctct ggtggtggtt ctggtggcgg ctctgagggt
     3241 ggcggctctg agggtggcgg ttctgagggt ggcggctctg agggtggcgg ttccggtggc
     3301 ggctccggtt ccggtgattt tgattatgaa aaaatggcaa acgctaataa gggggctatg
     3361 accgaaaatg ccgatgaaaa cgcgctacag tctgacgcta aaggcaaact tgattctgtc
     3421 gctactgatt acggtgctgc tatcgatggt ttcattggtg acgtttccgg ccttgctaat
     3481 ggtaatggtg ctactggtga ttttgctggc tctaattccc aaatggctca agtcggtgac
     3541 ggtgataatt cacctttaat gaataatttc cgtcaatatt taccttcttt gcctcagtcg
     3601 gttgaatgtc gcccttatgt ctttggcgct ggtaaaccat atgaattttc tattgattgt
     3661 gacaaaataa acttattccg tggtgtcttt gcgtttcttt tatatgttgc cacctttatg
     3721 tatgtatttt cgacgtttgc taacatactg cgtaataagg agtcttaagc tagctaacag
     3781 tcctatgaat caactactta gatggtatta gtgacctgta acagagcatt agcgcaaggt
     3841 gatttttgtc ttcttgcgct aattttttgt catcaaacct gtcgcactcc ttaatatttt
     3901 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3961 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4021 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4081 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4141 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4201 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4261 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4321 gctacagggc gcgtcaggtg
//
