LOCUS       sequence-343866-        6434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the SR6v15.d targeting the receptor binding 
            domain of SARS-CoV-2 spike protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6434)
  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 6434)
  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: pETx-SR6v15.d
FEATURES             Location/Qualifiers
     source          1..6434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             695..712
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(761..779)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      775..822
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(997..1002,1003..1019,1020..1025))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1056..1074)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1142..1246
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1247..1315,1316..2107)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1465..1484)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2278..2866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2767..2786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3020..3037
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3052..3191
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3117..3136)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3277..3299
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3293..3484)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4796..5878)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5840..5859
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5879..5956)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6162..6181
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6265..6284
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6265..6283
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6284..6308
                     /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)."
     RBS             6323..6345
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ctcttacata ttatggcgca tgggatggtt tcgtcaggca cctggtaagg aacgcgagtt
       61 cgtagccgcg atatcactgg acggtcagtt tacgaactac gcagactcgg tccgcggccg
      121 tttctcgata tctgcggaca gcgcaaaata tacggtctac ctgcagatga acagcctgaa
      181 accagaagat actgccgtct attactgcgc agcacttccg ttccttgatt actggggtca
      241 aggaactcaa gttaccgttt cttccggagg cggaggaagc ggaggaggag gctcgggagg
      301 aggcggctcc ggcggcggcg gttctggcgg aggaggttca gaggttcaac tggtagaatc
      361 aggaggcggt ttagttcagg cgggagatag cctgcgctta agctgcgcgg caagcggtga
      421 ctcttacata ttatggcgca tgggatggtt tcgtcaggca cctggtaagg aacgcgagtt
      481 cgtagccgcg atatcactgg acggtcagtt tacgaactac gcagactcgg tccgcggccg
      541 tttctcgata tctgcggaca gcgcaaaata tacggtctac ctgcagatga acagcctgaa
      601 accagaagat actgccgtct attactgcgc agcacttccg ttccttgatt actggggtca
      661 aggaactcaa gttaccgttt cttccggatc cggccatcac catcatcacc actaacggct
      721 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      781 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      841 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
      901 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
      961 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     1021 gtcaaacatg agaattaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     1081 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1141 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1201 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1261 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1321 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1381 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1441 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1501 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1561 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1621 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1681 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1741 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     1801 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1861 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1921 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1981 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2041 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2101 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2161 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2221 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2281 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2341 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2401 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2461 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2521 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2581 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2641 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2701 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2761 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2821 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2881 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2941 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3001 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     3061 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     3121 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3181 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3241 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3301 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3361 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     3421 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     3481 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     3541 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     3601 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     3661 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     3721 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     3781 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     3841 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     3901 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc
     3961 caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt
     4021 tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc
     4081 ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg
     4141 gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat
     4201 ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg
     4261 tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg
     4321 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
     4381 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
     4441 gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     4501 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     4561 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     4621 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     4681 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     4741 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     4801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4861 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     4921 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     4981 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     5041 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     5101 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     5161 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     5221 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     5281 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     5341 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     5401 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     5461 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     5521 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     5581 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     5641 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     5701 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     5761 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     5821 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     5881 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     5941 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     6001 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     6061 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     6121 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     6181 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     6241 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
     6301 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac catggaggtt
     6361 caactggtag aatcaggagg cggtttagtt caggcgggag atagcctgcg cttaagctgc
     6421 gcggcaagcg gtga
//
