LOCUS       sequence-345291-        6696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant fusion protein gene combining IgG-Fc binding domain Z (5
            repeats) of Staphylococcal Protein A and bioluminescence reporter 
            Nanoluciferase in pET28a bacterial expression plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6696)
  AUTHORS   Nandy S, Crum M, Wasden K, Strych U, Goyal A, Maranholkar V, Mo W, 
            Vu B, Kourentzi K, Willson RC
  TITLE     Protein A-nanoluciferase fusion protein for generalized, sensitive 
            detection of immunoglobulin G.
  JOURNAL   Anal Biochem. 2022 Oct 18:114929. doi: 10.1016/j.ab.2022.114929.
  PUBMED    36270332
REFERENCE   2  (bases 1 to 6696)
  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: "Anal 
            Biochem. 2022 Oct 18:114929. doi: 10.1016/j.ab.2022.114929."
COMMENT     Sequence Label: pSNRW-Z5-Nluc
FEATURES             Location/Qualifiers
     source          1..6696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..78)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     284..303
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     387..406
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        387..405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    406..430
                     /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             445..467
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             486..503
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             513..530
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1947..1964
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2017..2035)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2031..2078
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2115..2570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2202..2221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2412..2433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2662..3477)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2662..3477)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3385..3404
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3469..3488
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3599..4187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4088..4107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4341..4358
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4373..4512
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4438..4457)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4598..4620
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4614..4805)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5614..6696)
                     /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     6658..6677
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
       61 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
      121 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
      181 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
      241 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      301 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      361 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
      421 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggca
      481 gcagccatca tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatggtcg
      541 ataacaaatt caacaaagaa caacaaaacg cgttctatga gatcttacat ttacctaact
      601 taaacgaaga acaacgaaac gccttcatcc aaagtttaaa agatgaccca agccaaagcg
      661 ctaacctttt agcagaagct aaaaagctaa atgatgctca agcaccgaaa gtcgataaca
      721 aattcaacaa agaacaacaa aacgcgttct atgagatctt acatttacct aacttaaacg
      781 aagaacaacg aaacgccttc atccaaagtt taaaagatga cccaagccaa agcgctaacc
      841 ttttagcaga agctaaaaag ctaaatgatg ctcaagcacc gaaagtcgat aacaaattca
      901 acaaagaaca acaaaacgcg ttctatgaga tcttacattt acctaactta aacgaagaac
      961 aacgaaacgc cttcatccaa agtttaaaag atgacccaag ccaaagcgct aaccttttag
     1021 cagaagctaa aaagctaaat gatgctcaag caccgaaagt cgataacaaa ttcaacaaag
     1081 aacaacaaaa cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa
     1141 acgccttcat ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag
     1201 ctaaaaagct aaatgatgct caagcaccga aagtcgataa caaattcaac aaagaacaac
     1261 aaaacgcgtt ctatgagatc ttacatttac ctaacttaaa cgaagaacaa cgaaacgcct
     1321 tcatccaaag tttaaaagat gacccaagcc aaagcgctaa ccttttagca gaagctaaaa
     1381 agctaaatga tgctcaagca ccgaaagtcg acttacaagc tggcatggtt ttcaccctgg
     1441 aagacttcgt tggtgattgg cgtcagaccg caggttacaa tctggatcag gtcctggaac
     1501 agggcggtgt atctagcctg ttccagaacc tgggtgttag cgtcaccccg atccagcgta
     1561 tcgtactgag cggtgaaaac ggcctgaaaa tcgacatcca tgttatcatc ccgtacgagg
     1621 gtctgtctgg tgaccagatg ggtcagatcg aaaagatttt caaagtagtg tatccggttg
     1681 atgatcacca tttcaaagtt atcctgcact acggcaccct ggtaatcgac ggtgtgaccc
     1741 ctaacatgat tgactacttc ggtcgtcctt acgaaggcat cgcagttttc gatggcaaga
     1801 aaatcaccgt aaccggtacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1861 atccggacgg ctctctgctg ttccgtgtca ctatcaacgg cgttactggc tggcgtctgt
     1921 gcgagcgtat cctggcgtaa ctcgagcacc accaccacca ccactgagat ccggctgcta
     1981 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     2041 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     2101 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2161 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2221 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2281 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2341 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2401 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2461 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2521 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     2581 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2641 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     2701 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     2761 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     2821 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     2881 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     2941 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     3001 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     3061 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     3121 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     3181 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     3241 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     3301 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     3361 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     3421 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     3481 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     3541 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3601 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3661 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3721 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3781 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3841 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3901 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3961 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4021 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4081 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4141 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4201 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4261 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4321 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4381 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     4441 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     4501 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4561 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4621 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     4681 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     4741 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     4801 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     4861 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     4921 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     4981 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     5041 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     5101 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     5161 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     5221 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
     5281 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     5341 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     5401 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     5461 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     5521 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     5581 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5641 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5701 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     5761 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     5821 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     5881 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     5941 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     6001 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     6061 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     6121 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     6181 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     6241 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     6301 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     6361 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     6421 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     6481 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     6541 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     6601 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     6661 atactctgcg acatcgtata acgttactgg tttcac
//
