LOCUS       sequence-368406-        4971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express the Clover4-LUMABS-HIV sensor for Acquired 
            Immunodeficiency Virus antibody dection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4971)
  AUTHORS   Deng M, Yuan J, Yang H, Wu X, Wei X, Du Y, Wong G, Tao Y, Liu G, Jin
            Z, Chu J
  TITLE     A Genetically Encoded Bioluminescent System for Fast and Highly 
            Sensitive Detection of Antibodies with a Bright Green Fluorescent 
            Protein.
  JOURNAL   ACS Nano. 2021 Nov 2. doi: 10.1021/acsnano.1c05164.
  PUBMED    34726889
REFERENCE   2  (bases 1 to 4971)
  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: "ACS Nano. 
            2021 Nov 2. doi: 10.1021/acsnano.1c05164."
COMMENT     Sequence Label: pNCS-Clover4-LUMABS-HIV
FEATURES             Location/Qualifiers
     source          1..4971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             122..144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             164..181
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..217
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(221..229,230..244)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             263..280
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             290..307
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             428..439
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(563..565,566..568,569..1279)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFARYPDHMKLHDFFKAAMPEGYVQERTISFEDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKAKNGIK
                     ANFKIYHNVEDGSVQVADHYQQNTPIGDGPVLLPDNHYLSHQSTLSKDPNEKRDHMVLV
                     ERVTAAGITHGMDELYK"
     primer_bind     complement(608..629)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1736..2248
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2372..2390)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2386..2433
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2530..2985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2617..2636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2827..2848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3011..3115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3116..3184,3185..3976)
                     /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     complement(3334..3353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4147..4735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4636..4655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4889..4906
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaatcaccg agtttattct tgacacctga tgcgatgaat gatataatag gaaagtactg
      121 ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc atcatcatca
      181 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga
      241 taaggatccg atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg
      301 cggcagccat atggctagcg acgacaactt tatatacaag gcgaaggcgc tctatcccta
      361 tgatgcagat gatgatgacg catacgaaat ctcattcgaa caaaatgaaa tccttcaggt
      421 atcagacatt gaggggcggt ggtggaaagc ccgccgcgcc aatggagaga caggcatcat
      481 cccgtcgaat tatgttcagt tgattgacgg tcctgaggaa atgcatcggg gcggctcggg
      541 aggctcaggc agccatggat ccatggtgag caagggcgaa gagctgttca ccggggtggt
      601 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga
      661 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      721 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcgc
      781 caggtacccc gaccacatga agctgcacga cttcttcaag gccgccatgc ccgaaggcta
      841 cgtccaggag cgcaccatct ctttcgagga cgacggtacc tacaagaccc gcgccgaggt
      901 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      961 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat
     1021 catggccgac aaggccaaga acggcatcaa ggctaacttc aagatctacc acaacgttga
     1081 ggacggcagc gtgcaggtgg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1141 cgtgctgctg cccgacaacc actacctgag ccatcagtcc accctgagca aagaccccaa
     1201 cgagaagcgc gatcacatgg tcctggtgga gcgcgtgacc gccgccggga ttacacatgg
     1261 catggacgag ctgtacaagg aattctcggg cggagagcta gatcgctggg aaaaaatacg
     1321 ccttagaccg gggggttcgg gtggttcagg cggctcagga ggctccgggg gttccggagg
     1381 gagcggtgct gaagccgcag ccaaggaagc agcagctaaa gaggccgctg cgaaggaagc
     1441 tgccgcaaag gaggcggcgg cgaaagaggc ggcagcaaaa gccggatctg gtggcagtgg
     1501 tggctccggc gggtcaggtg gcagcggggg atcaggagct gaggcagccg ccaaagaggc
     1561 tgcggccaag gaggccgccg ctaaagaagc cgcggcaaaa gaggcagcgg caaaggaagc
     1621 ggctgcgaaa gccggaagtg gtgggtcggg cggctccggt ggctctggcg gcagtggcgg
     1681 tagtggcggg gaattagata ggtgggaaaa gatccggtta cgcccgggag gcagcatggt
     1741 cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca
     1801 agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc
     1861 gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat
     1921 cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt
     1981 gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga
     2041 cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt
     2101 cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga
     2161 gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg
     2221 ctggcggctg tgcgaacgca ttctggcgtc tagcggtgga ggatctattc gttcaaagcc
     2281 gctgccaccc ctacccgtga ccgggtaatg aattcgaagc ttgatccggc tgctaacaaa
     2341 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2401 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatct
     2461 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2521 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2581 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2641 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2701 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2761 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2821 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2881 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2941 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     3001 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3061 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3121 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3181 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3241 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3301 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3361 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3421 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3481 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3541 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3601 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3661 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3721 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3781 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3841 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3901 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3961 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4261 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4741 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4861 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4921 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca g
//
