LOCUS       sequence-404723-        4337 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 4337)
  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 4337)
  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_A8_His
FEATURES             Location/Qualifiers
     source          1..4337
                     /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             2469..2486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2509..2538
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      complement(3903..4331)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4013..4034)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4225..4244
                     /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 tgggggagga ttggtgcagg ctggaggctc tctgagactc tcctgtgcag
     2161 cctctggagg caccttcagt accgctgcca tgggctggtt ccgccaggct ccaggcgagg
     2221 agcgtgagtg tgtagcatct ataggctgga gaggtgttag aacatggtat gcagactccg
     2281 tgaagggccg atttaccatc tccagagata atccccagaa tacggtgtat ttgcagatga
     2341 acaacctgaa atctggcgac acggccgttt attactgcgc agcctcagtc ggcaactacg
     2401 ggttgccgtg ggcccacttc gaatatgact tctggggcca ggggatccag gtcaccgtct
     2461 cctcaggcca ccatcaccat caccattaat aaccaccatc atggcgcaga acaaaaactc
     2521 atctcagaag aggatctgtc ttaggccgaa actgttgaaa gttgtttagc aaaacctcat
     2581 acagaaaatt catttactaa cgtctggaaa gacgacaaaa ctttagatcg ttacgctaac
     2641 tatgagggct gtctgtggaa tgctacaggc gttgtggttt gtactggtga cgaaactcag
     2701 tgttacggta catgggttcc tattgggctt gctatccctg aaaatgaggg tggtggctct
     2761 gagggtggcg gttctgaggg tggcggttct gagggtggcg gtactaaacc tcctgagtac
     2821 ggtgatacac ctattccggg ctatacttat atcaaccctc tcgacggcac ttatccgcct
     2881 ggtactgagc aaaaccccgc taatcctaat ccttctcttg aggagtctca gcctcttaat
     2941 actttcatgt ttcagaataa taggttccga aataggcagg gtgcattaac tgtttatacg
     3001 ggcactgtta ctcaaggcac tgaccccgtt aaaacttatt accagtacac tcctgtatca
     3061 tcaaaagcca tgtatgacgc ttactggaac ggtaaattca gagactgcgc tttccattct
     3121 ggctttaatg aggatccatt cgtttgtgaa tatcaaggcc aatcgtctga cctgcctcaa
     3181 cctcctgtca atgctggcgg cggctctggt ggtggttctg gtggcggctc tgagggtggc
     3241 ggctctgagg gtggcggttc tgagggtggc ggctctgagg gtggcggttc cggtggcggc
     3301 tccggttccg gtgattttga ttatgaaaaa atggcaaacg ctaataaggg ggctatgacc
     3361 gaaaatgccg atgaaaacgc gctacagtct gacgctaaag gcaaacttga ttctgtcgct
     3421 actgattacg gtgctgctat cgatggtttc attggtgacg tttccggcct tgctaatggt
     3481 aatggtgcta ctggtgattt tgctggctct aattcccaaa tggctcaagt cggtgacggt
     3541 gataattcac ctttaatgaa taatttccgt caatatttac cttctttgcc tcagtcggtt
     3601 gaatgtcgcc cttatgtctt tggcgctggt aaaccatatg aattttctat tgattgtgac
     3661 aaaataaact tattccgtgg tgtctttgcg tttcttttat atgttgccac ctttatgtat
     3721 gtattttcga cgtttgctaa catactgcgt aataaggagt cttaagctag ctaacagtcc
     3781 tatgaatcaa ctacttagat ggtattagtg acctgtaaca gagcattagc gcaaggtgat
     3841 ttttgtcttc ttgcgctaat tttttgtcat caaacctgtc gcactcctta atattttgtt
     3901 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3961 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4021 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4081 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4141 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4201 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4261 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4321 acagggcgcg tcaggtg
//
