LOCUS       sequence-368273-        3256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template for generating CeVICA VHH/nanobody library.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3256)
  AUTHORS   Chen X, Gentili M, Hacohen N, Regev A
  TITLE     A cell-free nanobody engineering platform rapidly generates 
            SARS-CoV-2 neutralizing nanobodies.
  JOURNAL   Nat Commun. 2021 Sep 17;12(1):5506. doi: 10.1038/s41467-021-25777-z.
  PUBMED    34535642
REFERENCE   2  (bases 1 to 3256)
  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 
            Commun."; date: "2021-09-17"; volume: "12(1):5506. doi"; pages: " 
            10.1038/s41467-021-25777-z"
COMMENT     Sequence Label: pBx1-VHH template3-3XMyc-Spacer
FEATURES             Location/Qualifiers
     source          1..3256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     157..176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        157..175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             189..211
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             584..613
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             617..646
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             656..685
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(889..907)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      903..950
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1065..1186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1102..1121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1156..1175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1235..1251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1235..1251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1248..1270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1259..1275
                     /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)."
     promoter        complement(join(1283..1289,1290..1307,1308..1313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1328..1349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1466..1483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1637..2225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1717..1736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2396..3187,3188..3256))
                     /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     3019..3038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtcgacg gatcgggagt ttaaactaat acgactcact atagggtcta
      181 gaaataattt tgtttaactt taagaaggag atataccatg caggttcaac tgcaagaatc
      241 tggtggcggt ttagttcaag ccggtggtag ccttcgtctg agctgcgccg caagcggacg
      301 tacattcagc agctatgcaa tgggttggtt tcgccaggca cctggtaaag aacgtgaatt
      361 tgtggcagcc atttcaagca gtggtggcac ctactatgcc gattctgtta aaggtcgctt
      421 cactatttca cgcgataatg cgaagaatac cgtgtactta cagatgaact cactgaaacc
      481 tgaggatacc gccgtttatt attgtgcggc aggccgtgga ggcggtagcg actattgggg
      541 tcagggcaca caagtcacgg tctcaagcgg cggaagctct ggtgaacaga aactgatttc
      601 agaagaggat ctggcagaac agaaattaat ctcagaggaa gacctgggtt ctggtgagca
      661 aaagttaatt tctgaagaag acctggcagg tggctctggt gatggaggta gctctggcga
      721 tggtgcggat ggcagcggtg gtgcatctgg ttcaagcgct tacagcgatg atgttagtgc
      781 aagcggtcgt acctaccgca tcttctcata taactacgcc agctatagcg actgggttta
      841 aaccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      901 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      961 gaactatatc cggtgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     1021 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     1081 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1141 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     1201 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1261 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1321 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1381 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1441 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1501 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1561 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2041 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2161 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2221 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2281 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2341 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2401 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2461 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2521 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2581 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2641 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2701 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2761 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2821 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2881 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2941 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3001 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3061 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3121 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3181 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3241 aatgttgaat actcat
//
