LOCUS       sequence-413320-        4334 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of nanobody.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4334)
  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 4334)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_B5-5_His
FEATURES             Location/Qualifiers
     source          1..4334
                     /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             2466..2483
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2506..2535
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      complement(3900..4328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4010..4031)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4222..4241
                     /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 caggtccagt
     2101 tggtagaaag tggcggtggt ttggtgcaac ccggcggctc gttgcgttta tcctgcgccg
     2161 tttttgggtc cactctggat aattatagcg tgggctggtt tcgccagtta ccagggaaag
     2221 agcgcgaggg attggcctgc ttctcacgcc gctatggcgc cccatattat gcggactctg
     2281 ccaaagatcg tttcactatt tcccgcgata acgctaagaa tacggttgac ttacagctga
     2341 acaacttaaa acccgaggat acggcagtct attattgtgc gacacgttct ggaccctatt
     2401 gcaccacgag cgtatcggat tttgatgatt gggggcgtgg gactcaagta accgtgtcct
     2461 cgggccacca tcaccatcac cattaataac caccatcatg gcgcagaaca aaaactcatc
     2521 tcagaagagg atctgtctta ggccgaaact gttgaaagtt gtttagcaaa acctcataca
     2581 gaaaattcat ttactaacgt ctggaaagac gacaaaactt tagatcgtta cgctaactat
     2641 gagggctgtc tgtggaatgc tacaggcgtt gtggtttgta ctggtgacga aactcagtgt
     2701 tacggtacat gggttcctat tgggcttgct atccctgaaa atgagggtgg tggctctgag
     2761 ggtggcggtt ctgagggtgg cggttctgag ggtggcggta ctaaacctcc tgagtacggt
     2821 gatacaccta ttccgggcta tacttatatc aaccctctcg acggcactta tccgcctggt
     2881 actgagcaaa accccgctaa tcctaatcct tctcttgagg agtctcagcc tcttaatact
     2941 ttcatgtttc agaataatag gttccgaaat aggcagggtg cattaactgt ttatacgggc
     3001 actgttactc aaggcactga ccccgttaaa acttattacc agtacactcc tgtatcatca
     3061 aaagccatgt atgacgctta ctggaacggt aaattcagag actgcgcttt ccattctggc
     3121 tttaatgagg atccattcgt ttgtgaatat caaggccaat cgtctgacct gcctcaacct
     3181 cctgtcaatg ctggcggcgg ctctggtggt ggttctggtg gcggctctga gggtggcggc
     3241 tctgagggtg gcggttctga gggtggcggc tctgagggtg gcggttccgg tggcggctcc
     3301 ggttccggtg attttgatta tgaaaaaatg gcaaacgcta ataagggggc tatgaccgaa
     3361 aatgccgatg aaaacgcgct acagtctgac gctaaaggca aacttgattc tgtcgctact
     3421 gattacggtg ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat
     3481 ggtgctactg gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat
     3541 aattcacctt taatgaataa tttccgtcaa tatttacctt ctttgcctca gtcggttgaa
     3601 tgtcgccctt atgtctttgg cgctggtaaa ccatatgaat tttctattga ttgtgacaaa
     3661 ataaacttat tccgtggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta
     3721 ttttcgacgt ttgctaacat actgcgtaat aaggagtctt aagctagcta acagtcctat
     3781 gaatcaacta cttagatggt attagtgacc tgtaacagag cattagcgca aggtgatttt
     3841 tgtcttcttg cgctaatttt ttgtcatcaa acctgtcgca ctccttaata ttttgttaaa
     3901 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3961 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4021 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4081 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4141 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4201 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4261 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4321 gggcgcgtca ggtg
//
