LOCUS       sequence-337035-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 RBD in bacterial system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Ahmad J, Jiang J, Boyd LF, Natarajan K, Margulies DH
  TITLE     Synthetic nanobody-SARS-CoV-2 receptor-binding domain structures 
            identify distinct epitopes.
  JOURNAL   bioRxiv. 2021 Jan 27. doi: 10.1101/2021.01.27.428466.
  PUBMED    33532775
REFERENCE   2  (bases 1 to 5993)
  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: "bioRxiv. 
            2021 Jan 27. doi: 10.1101/2021.01.27.428466."
COMMENT     Sequence Label: pET21b-RBD-JA1
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     937..956
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(976..1053)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1259..1278
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1362..1381
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1362..1380
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1381..1405
                     /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             1420..1442
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2086..2103
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2156..2174)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2170..2217
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2254..2709
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2341..2360)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2551..2572
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2735..2839
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2840..2908,2909..3700)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3058..3077)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3871..4459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4360..4379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4613..4630
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4645..4784
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4710..4729)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4870..4892
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4886..5077)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5886..5993,1..975))
                     /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"
ORIGIN
        1 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
       61 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
      121 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
      181 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
      241 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
      301 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
      361 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
      421 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
      481 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
      541 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
      601 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
      661 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
      721 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
      781 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
      841 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
      901 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
      961 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     1021 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     1081 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     1141 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     1201 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     1261 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     1321 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg
     1381 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
     1441 gatatacata tgaatctttg tccgttcggt gaggttttta acgcgacaag gttcgctagt
     1501 gtatatgctt ggaaccgaaa gagaatctcc aattgcgtag ctgattactc cgttctctat
     1561 aacagtgcgt ccttttcaac ctttaagtgt tacggcgttt ctccaacgaa gctgaatgat
     1621 ctctgtttta cgaacgtgta tgctgactct ttcgttatac ggggggacga agtgagacag
     1681 atagcaccag gtcagactgg gaagatagcg gattacaact ataagttgcc cgatgatttt
     1741 acggggtgcg taatcgcatg gaactcaaac aacctcgact ccaaagtagg tggtaattat
     1801 aattacttgt atcgcctgtt tcgaaagagc aatttgaagc cttttgagcg ggatatttca
     1861 accgaaattt accaagcagg cagtacgcca tgtaacggag tagagggatt taattgctac
     1921 tttcctcttc aatcttatgg ctttcaacca acaaacggag tggggtatca accttataga
     1981 gtggtagtat tgtcctttga gctcctccac gccccggcta cagtttgtgg gcccaaaaag
     2041 ggtgagaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca ccaccaccac
     2101 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2161 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2221 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2281 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2341 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2401 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2461 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2521 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2581 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2641 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     2701 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2761 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2821 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2881 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2941 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3001 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3061 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3121 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3181 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3241 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3301 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3361 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3421 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3481 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3541 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3601 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3661 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3721 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3781 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3841 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3901 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3961 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4021 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4081 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4141 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4201 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4261 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4321 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4381 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4441 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4501 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4561 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4621 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4681 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4741 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     4801 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4861 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4921 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     4981 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     5041 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     5101 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     5161 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     5221 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     5281 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     5341 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     5401 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     5461 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     5521 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     5581 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     5641 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     5701 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     5761 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     5821 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     5881 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5941 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tct
//
