LOCUS       sequence-368403-        8001 bp    DNA     circular SYN 15-APR-2025
DEFINITION  To express the Clover4-LUMABS-S1 RBD sensor for SARS-CoV-2 antibody 
            dection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8001)
  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 8001)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pFastBac1-Clover4-LUMABS-S1 RBD
FEATURES             Location/Qualifiers
     source          1..8001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3505..3510,3511..3527,3528..3533))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3885..3976
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3901..3918
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     sig_peptide     4110..4169
                     /codon_start=1
                     /gene="Autographa californica gp64"
                     /product="signal sequence from the baculovirus envelope 
                     glycoprotein GP64"
                     /label=GP64 signal sequence
                     /translation="MVSAIVLYVLLAAAAHSAFA"
     CDS             4302..4313
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(4437..4439,4440..4442,4443..5153)
                     /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(4482..4503)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             6738..7250
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             7323..7340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    7491..7625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7528..7547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7582..7601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7654..7819
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     3361 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3421 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3481 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3541 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3601 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3661 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3721 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3781 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3841 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     3901 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     3961 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatcccg
     4021 gtccgaagcg cgcggaattc aaggcctacg tcgacatgct actagtaaat cagtcacacc
     4081 aaggcttcaa taaggaacac acaagcaaga tggtaagcgc tattgtttta tatgtgcttt
     4141 tggcggcggc ggcgcattct gcctttgcgg gtggatctgg gggcactgct agcgacgaca
     4201 actttatata caaggcgaag gcgctctatc cctatgatgc agatgatgat gacgcatacg
     4261 aaatctcatt cgaacaaaat gaaatccttc aggtatcaga cattgagggg cggtggtgga
     4321 aagcccgccg cgccaatgga gagacaggca tcatcccgtc gaattatgtt cagttgattg
     4381 acggtcctga ggaaatgcat cggggcggct cgggaggctc aggcagccat ggatccatgg
     4441 tgagcaaggg cgaagagctg ttcaccgggg tggtgccaat cctggtcgag ctggacggcg
     4501 acgtaaacgg ccacaagttc agcgtccgcg gcgagggcga gggcgatgcc accaacggca
     4561 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     4621 tgaccacctt cggctacggc gtggcctgct tcgccaggta ccccgaccac atgaagctgc
     4681 acgacttctt caaggccgcc atgcccgaag gctacgtcca ggagcgcacc atctctttcg
     4741 aggacgacgg tacctacaag acccgcgccg aggtgaagtt cgagggtgac accctggtga
     4801 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     4861 tggagtacaa cttcaacagc cacaacgtct atatcatggc cgacaaggcc aagaacggca
     4921 tcaaggctaa cttcaagatc taccacaacg ttgaggacgg cagcgtgcag gtggccgacc
     4981 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     5041 tgagccatca gtccaccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgg
     5101 tggagcgcgt gaccgccgcc gggattacac atggcatgga cgagctgtac aaggaattct
     5161 cgggcggaaa tattacaaac ttgtgccctt ttggtgaagt ttttaacgcc accagatttg
     5221 catctgttta tgcttggaac aggaagagaa tcagcaactg tgttgctgat tattctgtcc
     5281 tatataattc cgcatcattt tccactttta agtgttatgg agtgtctcct actaaattaa
     5341 atgatctctg ctttactaat gtctatgcag attcatttgt aattagaggt gatgaagtca
     5401 gacaaatcgc tccagggcaa actggaaaga ttgctgatta taattataaa ttaccagatg
     5461 attttacagg ctgcgttata gcttggaatt ctaacaatct tgattctaag gttggtggta
     5521 attataatta cctgtataga ttgtttagga agtctaatct caaacctttt gagagagata
     5581 tttcaactga aatctatcag gccggtagca caccttgtaa tggtgttgaa ggttttaatt
     5641 gttactttcc tttacaatca tatggtttcc aacccactaa tggtgttggt taccaaccat
     5701 acagagtagt agtactttct tttgaacttc tacatgcacc agcaactgtt tgtggaccta
     5761 aaaagtctgg gggttcgggt ggttcaggcg gctcaggagg ctccgggggt tccggaggga
     5821 gcggtgctga agccgcagcc aaggaagcag cagctaaaga ggccgctgcg aaggaagctg
     5881 ccgcaaagga ggcggcggcg aaagaggcgg cagcaaaagc cggatctggt ggcagtggtg
     5941 gctccggcgg gtcaggtggc agcgggggat caggagctga ggcagccgcc aaagaggctg
     6001 cggccaagga ggccgccgct aaagaagccg cggcaaaaga ggcagcggca aaggaagcgg
     6061 ctgcgaaagc cggaagtggt gggtcgggcg gctccggtgg ctctggcggc agtggcggta
     6121 gtggcgggaa tattacaaac ttgtgccctt ttggtgaagt ttttaacgcc accagatttg
     6181 catctgttta tgcttggaac aggaagagaa tcagcaactg tgttgctgat tattctgtcc
     6241 tatataattc cgcatcattt tccactttta agtgttatgg agtgtctcct actaaattaa
     6301 atgatctctg ctttactaat gtctatgcag attcatttgt aattagaggt gatgaagtca
     6361 gacaaatcgc tccagggcaa actggaaaga ttgctgatta taattataaa ttaccagatg
     6421 attttacagg ctgcgttata gcttggaatt ctaacaatct tgattctaag gttggtggta
     6481 attataatta cctgtataga ttgtttagga agtctaatct caaacctttt gagagagata
     6541 tttcaactga aatctatcag gccggtagca caccttgtaa tggtgttgaa ggttttaatt
     6601 gttactttcc tttacaatca tatggtttcc aacccactaa tggtgttggt taccaaccat
     6661 acagagtagt agtactttct tttgaacttc tacatgcacc agcaactgtt tgtggaccta
     6721 aaaagtctgg aggcagcatg gtcttcacac tcgaagattt cgttggggac tggcgacaga
     6781 cagccggcta caacctggac caagtccttg aacagggagg tgtgtccagt ttgtttcaga
     6841 atctcggggt gtccgtaact ccgatccaaa ggattgtcct gagcggtgaa aatgggctga
     6901 agatcgacat ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga
     6961 tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc
     7021 actatggcac actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc
     7081 cgtatgaagg catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga
     7141 acggcaacaa aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag
     7201 taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg cattctggcg tctagcggtg
     7261 gaggatctat tcgttcaaag ccgctgccac ccctacccgt gaccggggga ggctctacgc
     7321 gtcatcacca tcaccatcac taactcgagg catgcggtac caagcttgtc gagaagtact
     7381 agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     7441 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     7501 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     7561 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     7621 ggatctgatc actgcttgag cctaggagat ccgaaccaga taagtgaaat ctagttccaa
     7681 actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta
     7741 ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca
     7801 actattttgt ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc
     7861 ctgccaactc catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa
     7921 aatttttctg tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt
     7981 tgcagcctga atggcgaatg g
//
