LOCUS       sequence-316777-        6037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for RBD of bat coronavirus isolate 
            RsSHC014.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6037)
  AUTHORS   Chan KK, Tan TJC, Narayanan KK, Procko E
  TITLE     An engineered decoy receptor for SARS-CoV-2 broadly binds protein S 
            sequence variants.
  JOURNAL   Sci Adv. 2021 Feb 17;7(8). pii: 7/8/eabf1738. doi: 
            10.1126/sciadv.abf1738. Print 2021 Feb.
  PUBMED    33597251
REFERENCE   2  (bases 1 to 6037)
  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: "Sci Adv."; 
            date: "2021-02-17"; volume: "7(8). pii: 7/8/eabf1738. doi"; pages: "
            10.1126/sciadv.abf1738. Print 2021 Feb"
COMMENT     Sequence Label: pcDNA3-RsSHC014-RBD-8h
FEATURES             Location/Qualifiers
     source          1..6037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        508..711
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     662..682
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     756..775
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        756..774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1454..1477
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(1521..1538)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1527..1751
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1797..2225
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1884..1903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2094..2115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2234..2254)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2239..2568
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2419..2554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2481..2500
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2635..3429
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2689..2708)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3299..3318
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3603..3724
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3640..3659)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3694..3713
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3773..3789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3773..3789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3786..3808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3797..3813
                     /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)."
     promoter        complement(join(3821..3827,3828..3845,3846..3851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3866..3887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4004..4021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4175..4763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4255..4274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4934..5725,5726..5794))
                     /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     5557..5576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5795..5899)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5974..5993)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
       61 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      121 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      181 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      301 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      361 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      421 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      481 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      541 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      601 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      661 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      721 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      781 caagctggct agcgccacca tgaagaccat catcgccctg agctacatct tctgcctggt
      841 gttcgccacg aatttgtgcc catttgggga ggtatttaac gcaactacat tcccttctgt
      901 ttacgcgtgg gaaaggaaga gaatcagtaa ttgcgttgct gattattctg tgctttacaa
      961 ctctacttct ttcagtacat tcaagtgcta cggagtttca gcaactaaac tcaacgactt
     1021 gtgcttctca aatgtgtacg ctgatagctt cgtggtcaag ggtgacgatg tccgccaaat
     1081 agcgccaggc caaactgggg tgattgcgga ttacaattat aaactgcccg acgatttctt
     1141 gggttgtgtg cttgcctgga acactaattc aaaggattca tccacttccg gcaactacaa
     1201 ttacttgtat agatgggtcc gacggtcaaa attgaatcca tacgaaaggg atctgtctaa
     1261 tgacatctat tccccaggag gccagagctg tagcgccgtt gggcctaact gctataatcc
     1321 actgcgaccg tatggctttt tcactacagc tggggtagga catcagccgt atagggttgt
     1381 agtcttgtct tttgaattgc ttaatgcgcc agccactgtc tgtggaccta agctgagcac
     1441 tgacggatcc ggtcaccatc accaccacca tcatcattga taactcgagt ctagagggcc
     1501 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1561 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1621 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1681 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1741 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1801 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1861 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1921 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1981 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2041 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2101 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2161 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2221 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2341 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2401 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2461 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2521 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2581 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2641 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2701 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2761 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     2821 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2881 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2941 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3001 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3061 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3121 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3181 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3241 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3301 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3361 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3421 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3481 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3541 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3601 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3661 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3721 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3901 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3961 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4021 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4081 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4141 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4201 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4261 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4321 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4381 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4441 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4501 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4561 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4621 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4681 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4741 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4801 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4861 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4921 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4981 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5041 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5101 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5161 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5221 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5281 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5341 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5401 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5461 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5521 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5581 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5641 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5701 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5761 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5821 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5881 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     5941 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6001 aagccagtat ctgctccctg cttgtgtgtt ggaggtc
//
