LOCUS       sequence-345289-        6348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant fusion protein gene combining IgG-Fc binding domain Z (3
            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 6348)
  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 6348)
  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-Z3-Nluc
FEATURES             Location/Qualifiers
     source          1..6348
                     /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             1599..1616
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1669..1687)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1683..1730
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1767..2222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1854..1873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2064..2085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2314..3129)
                     /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(2314..3129)
                     /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     3037..3056
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3121..3140
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3251..3839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3740..3759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3993..4010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4025..4164
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4090..4109)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4250..4272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4266..4457)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5266..6348)
                     /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     6310..6329
                     /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 cgacttacaa gctggcatgg
     1081 ttttcaccct ggaagacttc gttggtgatt ggcgtcagac cgcaggttac aatctggatc
     1141 aggtcctgga acagggcggt gtatctagcc tgttccagaa cctgggtgtt agcgtcaccc
     1201 cgatccagcg tatcgtactg agcggtgaaa acggcctgaa aatcgacatc catgttatca
     1261 tcccgtacga gggtctgtct ggtgaccaga tgggtcagat cgaaaagatt ttcaaagtag
     1321 tgtatccggt tgatgatcac catttcaaag ttatcctgca ctacggcacc ctggtaatcg
     1381 acggtgtgac ccctaacatg attgactact tcggtcgtcc ttacgaaggc atcgcagttt
     1441 tcgatggcaa gaaaatcacc gtaaccggta ccctgtggaa cggcaacaaa attatcgacg
     1501 agcgcctgat caatccggac ggctctctgc tgttccgtgt cactatcaac ggcgttactg
     1561 gctggcgtct gtgcgagcgt atcctggcgt aactcgagca ccaccaccac caccactgag
     1621 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     1681 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     1741 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     1801 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1861 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1921 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1981 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2041 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2101 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2161 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     2221 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2281 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     2341 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     2401 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     2461 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     2521 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     2581 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     2641 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     2701 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2761 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2821 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2881 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2941 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     3001 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     3061 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     3121 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     3181 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3361 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3841 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3901 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3961 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4021 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     4081 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     4141 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     4201 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4261 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     4321 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     4381 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     4441 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     4501 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     4561 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     4621 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     4681 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     4741 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     4801 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     4861 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     4921 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     4981 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     5041 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     5101 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     5161 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     5221 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     5281 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5341 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     5401 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5461 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     5521 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     5581 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     5641 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     5701 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5761 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     5821 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     5881 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5941 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     6001 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     6061 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     6121 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     6181 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     6241 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     6301 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcac
//
