LOCUS       sequence-316758-        6037 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression plasmid for RBD of bat coronavirus isolate 
            LYRa11.
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 Apr 14, 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-LYRa11-RBD-8h
FEATURES             Location/Qualifiers
     source          1..6037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        166..545
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        546..749
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     700..720
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     794..813
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        794..812
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1492..1515
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(1559..1576)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1565..1789
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1835..2263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1922..1941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2132..2153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2272..2292)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2277..2606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2457..2592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2519..2538
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2673..3467
                     /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(2727..2746)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3337..3356
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3641..3762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3678..3697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3732..3751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3811..3827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3811..3827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3824..3846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3835..3851
                     /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(3859..3865,3866..3883,3884..3889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3904..3925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4042..4059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4213..4801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4293..4312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4972..5763,5764..5832))
                     /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     5595..5614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5833..5937)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6012..6031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
       61 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
      121 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
      181 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      241 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      301 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      361 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      421 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      481 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      541 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      601 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      661 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      721 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
      781 ggcttatcga aattaatacg actcactata gggagaccca agctggctag cgccaccatg
      841 aagaccatca tcgccctgag ctacatcttc tgcctggtgt tcgccaccaa cttgtgtccc
      901 ttcggcgagg tgtttaacgc aactactttc ccatctgtat atgcgtggga gaggaagaga
      961 atttctaatt gtgtagccga ttatagcgtt ctctacaact ccacgagttt cagcacgttc
     1021 aagtgctatg gagtgtcagc gatcaaactg aacgacctct gtttttctaa cgtttacgca
     1081 gatagctttg tagtgaaggg agatgatgta cggcaaattg cgccaggtca aacaggagtg
     1141 attgcagatt ataactacaa attgcctgac gattttatgg gctgtgtctt ggcatggaat
     1201 acaagaaaca tagatgccac ttcttcaggt aattttaact acaagtatag aagtttgcgc
     1261 cacgggaagt tgagaccctt cgagagagac atcagcaatg ttccattctc cccggacgga
     1321 aaaccatgta ctcctccggc tttcaattgt tactggccct tgaatgatta tgggttttat
     1381 acgaccaacg gaattgggta tcaaccatat cgcgtcgttg tcctgtcatt tgaactgctc
     1441 aatgcgcctg ctacagtctg cggacccaag ctctctaccg atggatccgg tcaccatcac
     1501 caccaccatc atcattgata actcgagtct agagggcccg tttaaacccg ctgatcagcc
     1561 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     1621 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     1681 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     1741 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     1801 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     1861 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1921 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1981 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2041 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2101 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2161 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2221 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     2281 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2341 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2401 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2461 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2521 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     2581 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     2641 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     2701 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2761 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2821 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     2881 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2941 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3001 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3061 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3121 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3181 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3241 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3301 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3361 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3421 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3481 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     3541 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     3601 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     3661 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3721 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     3781 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3841 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3901 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3961 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4021 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4081 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4141 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4621 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4861 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4921 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4981 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5041 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5101 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5161 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5221 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5281 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5341 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5401 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5461 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5521 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5581 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5641 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5701 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5761 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5821 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5881 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5941 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     6001 tgcactctca gtacaatctg ctctgatgcc gcatagt
//
