LOCUS       sequence-316809-        7136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transposon-based SARS-CoV-2 RBD stable expression vector for protein
            production .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7136)
  AUTHORS   Arias-Arias JL, Molina-Castro SE, Monturiol-Gross L, Lomonte B, 
            Corrales-Aguilar E
  TITLE     Stable production of recombinant SARS-CoV-2 receptor-binding domain 
            in mammalian cells with co-expression of a fluorescent reporter and 
            its validation as antigenic target for COVID-19 serology testing.
  JOURNAL   Biotechnol Rep (Amst). 2023 Mar;37:e00780. doi: 
            10.1016/j.btre.2022.e00780. Epub 2022 Dec 31.
  PUBMED    36619904
REFERENCE   2  (bases 1 to 7136)
  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: "Biotechnol 
            Rep (Amst)."; date: "2023-03"; volume: "37:e00780. doi"; pages: " 
            10.1016/j.btre.2022.e00780. Epub 2022 Dec 31"
COMMENT     Sequence Label: pSBbi-RP-CoV2/RBD-puro
FEATURES             Location/Qualifiers
     source          1..7136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1208..1217
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1928..1945
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1968..1985)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1974..2198
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2630..2734
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2735..2803,2804..3595)
                     /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(2953..2972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3766..4354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4255..4274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4508..4525
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4899..4918
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4917..4972
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     5018..5037
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(5137..5736)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5220..5240)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5717..5736
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(5737..5793)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(5803..6504)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(5886..5909)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(6084..6103)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6353..6371
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        join(7136,1..1182)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccatactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt aaggtgtcgt gaaaactacc ccaagctggc
     1201 ctctgaggcc accatgggct ttgtgtttct ggtgctgctg cctctggtca gttctcagag
     1261 agtgcagccc accgagagca tcgtgcggtt ccctaacatc acaaacctct gccccttcgg
     1321 cgaggtgttc aacgccacca gattcgcctc tgtgtacgcc tggaatagaa aaagaatcag
     1381 caattgcgtg gccgactaca gcgtgctgta taacagcgcc agcttcagca ccttcaagtg
     1441 ctacggcgtg tctccaacca agctgaacga cctgtgcttc acaaatgtgt acgccgacag
     1501 cttcgtgatt agaggagatg aggtgcggca gatcgctcct ggccagaccg gaaagatcgc
     1561 cgattacaac tataagctgc ccgacgactt caccggctgt gttatcgctt ggaacagcaa
     1621 caacctggac tctaaggtgg gcggaaatta caactacctg tacagactgt tccgcaaaag
     1681 caacctgaag cctttcgaaa gagacatcag caccgaaatc taccaggccg gcagcacacc
     1741 ttgtaatggc gtggaaggct ttaactgcta cttccccctg cagagctacg gcttccagcc
     1801 aacaaacggc gttggctacc aaccttaccg ggtggtggtc ctgagctttg agctgctgca
     1861 cgcccctgct accgtgtgcg gccctaagaa gtccacaaac ctggtgaaga acaaatgtgt
     1921 gaacttccac caccatcacc accactgatc tagagctcgc tgatcagcct cgactgtgcc
     1981 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2041 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2101 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2161 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     2221 ctgggcctac tagatagcgg accccttacc gaaacatcgc cgcattctgc agaggagtcg
     2281 aacttagtgt atgtaaactt ctgacccact ggaattgtga tacagtgaat tataagtgaa
     2341 ataatctgtc tgtaaacaat tgttggaaaa atgacttgtg tcatgcacaa agtagatgtc
     2401 ctaactgact tgccaaaact attgtttgtt aacaagaaat ttgtggagta gttgaaaaac
     2461 gagttttaat gactccaact taagtgtatg taaacttccg acttcaactg tatagggatc
     2521 caattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     2581 ctgtgcggta tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2641 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2701 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2761 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2821 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2881 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2941 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3001 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3061 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3121 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3181 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3241 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3301 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3361 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3421 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3481 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3541 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3601 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3661 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3721 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3781 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3841 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3901 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3961 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4021 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4081 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4141 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4201 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4261 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4321 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4381 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4441 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4501 cgagcgcagc gagtcagtga gcgaggaagc ggatccctat acagttgaag tcggaagttt
     4561 acatacacct tagccaaata catttaaact cactttttca caattcctga catttaatcc
     4621 tagtaaaaat tccctgtctt aggtcagtta ggatcaccac tttattttaa gaatgtgaaa
     4681 tatcagaata atagtagaga gaatgattca tttcagcttt tatttctttc atcacattcc
     4741 cagtgggtca gaagtttaca tacactcgac tcctctgcag aatgcggcga tgtttcggta
     4801 aggggtcccg agtagctagt tcatggcagc cagcatatgg catatgttgc caaactctaa
     4861 accaaatact cattctgatg ttttaaatga tttgccctcc catatgtcct tccgagtgag
     4921 agacacaaaa aattccaaca cactattgca atgaaaataa atttccttta ttagccagaa
     4981 gtcagatgct caaggggctt catgatgtcc ccataatttt tggcagaggg aaaaagatct
     5041 cagtggtatt tgtgagccag ggcattggcc acaccagcca ccaccttctg ataggcagcc
     5101 tgcacctgag gagtgaattc acgacaggcc ttcgaatcag gcaccgggct tgcgggtcat
     5161 gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga agccgagccg
     5221 ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg
     5281 ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc
     5341 gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag
     5401 gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg
     5461 gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac
     5521 cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa
     5581 gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt
     5641 ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg
     5701 ggtggcgagg cgcaccgtgg gcttgtactc ggtcatgggg ccaggattct cctcgacgtc
     5761 accagcctgc ttcagcaggc tgaagttagt agctccactg cccttgtaca gctcgtccat
     5821 gccgtacagg aacaggtggt ggcggccctc ggagcgctcg tactgttcca cgatggtgta
     5881 gtcctcgttg tgggaggtga tgtccagctt ggtgtccacg tagtagtagc cgggcagttg
     5941 cacgggcttc ttggccatgt agatggtctt gaactccacc aggtagtggc cgccgtcctt
     6001 cagcttcagg gcctggtgga tctcgccctt cagcacgccg tcgcgggggt acaggcgctc
     6061 ggtggaggcc tcccagccca ttgtcttctt ctgcattacg gggccgtcgg gggggaagtt
     6121 ggtgccgcgc atcttcacct tgtagatcag cgtgccgtcc tgcagggagg agtcctgggt
     6181 cacggtcacc agaccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg
     6241 gaaggacagc ttcttgtaat cggggatgtc ggcggggtgc ttcacgtacg ccttggagcc
     6301 gtacatgaac tggggggaca ggatgtccca ggcgaagggc agggggccgc ccttggtcac
     6361 cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc
     6421 gaactcatgg ccgttcatgg agccctccat gcgcaccttg aagcgcatga actctttgat
     6481 gacctcctcg cccttgctca acatggtggc ctcaggtgca gaggtttcta cagggaaaga
     6541 gagaaaggct acagttagtt tgctgtgatg atgctaagtg ccgaggtcag atgtctatgg
     6601 attgtacacg gaatcccaac gcaggaacca aaaaacactc acccaaaaaa atggcgctaa
     6661 ggccgagaga aaggcgaagt tccgtctacg gctatatacc ttgaaacagg ggcggcagaa
     6721 gtgacgcaac gcggttgcgc ggacggcttc ctgtgttttc attggtcgaa gcgttctgcg
     6781 cgtgcgcaga gattccccag ggaaggcggg ccctcttccg aatgtccaat ccggacattc
     6841 ttccggttgg acctccgcgg tcagcgctgt gcccggcgag agatcacgtg gggcgcggag
     6901 gcggtgctgc tggggcacgg ccgtccagcc tcggcggcca tatttttgag gggctgttca
     6961 tctcgttcac acgctctgtc cgccatgttt gtgagtggaa gcgccattac cccttcaagc
     7021 gactgaaggc tgcagggcct ctggtggccc gcatggggag accagacccg ccaggcccgc
     7081 ctttccgcac tcagtccggg cttactttat tttgtgagac agggtctcgc ctagag
//
