LOCUS       sequence-368409-        4953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express the Clover4-LUMABS-HA sensor for Avian Influenza Virus 
            antibody detection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4953)
  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 4953)
  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-HA
FEATURES             Location/Qualifiers
     source          1..4953
                     /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             1295..1321
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1682..1708
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1718..2230
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2354..2372)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2368..2415
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2512..2967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2599..2618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2809..2830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2993..3097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3098..3166,3167..3958)
                     /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(3316..3335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4129..4717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4618..4637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4871..4888
                     /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 cggatatccg tacgatgtgc cggattacgc
     1321 ggggggttcg ggtggttcag gcggctcagg aggctccggg ggttccggag ggagcggtgc
     1381 tgaagccgca gccaaggaag cagcagctaa agaggccgct gcgaaggaag ctgccgcaaa
     1441 ggaggcggcg gcgaaagagg cggcagcaaa agccggatct ggtggcagtg gtggctccgg
     1501 cgggtcaggt ggcagcgggg gatcaggagc tgaggcagcc gccaaagagg ctgcggccaa
     1561 ggaggccgcc gctaaagaag ccgcggcaaa agaggcagcg gcaaaggaag cggctgcgaa
     1621 agccggaagt ggtgggtcgg gcggctccgg tggctctggc ggcagtggcg gtagtggcgg
     1681 gtacccatat gacgtcccag actatgccgg aggcagcatg gtcttcacac tcgaagattt
     1741 cgttggggac tggcgacaga cagccggcta caacctggac caagtccttg aacagggagg
     1801 tgtgtccagt ttgtttcaga atctcggggt gtccgtaact ccgatccaaa ggattgtcct
     1861 gagcggtgaa aatgggctga agatcgacat ccatgtcatc atcccgtatg aaggtctgag
     1921 cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca
     1981 tcactttaag gtgatcctgc actatggcac actggtaatc gacggggtta cgccgaacat
     2041 gatcgactat ttcggacggc cgtatgaagg catcgccgtg ttcgacggca aaaagatcac
     2101 tgtaacaggg accctgtgga acggcaacaa aattatcgac gagcgcctga tcaaccccga
     2161 cggctccctg ctgttccgag taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg
     2221 cattctggcg tctagcggtg gaggatctat tcgttcaaag ccgctgccac ccctacccgt
     2281 gaccgggtaa tgaattcgaa gcttgatccg gctgctaaca aagcccgaaa ggaagctgag
     2341 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2401 ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat agcgaagagg
     2461 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     2521 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2581 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2641 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2701 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2761 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2821 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2881 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2941 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     3001 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3061 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3121 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3181 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3241 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3301 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3361 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3421 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3481 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3541 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3601 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3661 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3721 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3781 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3841 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3901 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3961 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4021 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4081 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4141 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4201 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4261 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4321 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4381 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4441 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4501 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4561 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4621 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4681 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4741 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4801 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4861 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4921 tctccccgcg cgttggccga ttcattaatg cag
//
