LOCUS       sequence-322928-        6031 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression plasmid for RBD of SARS-CoV-2 (N501Y mutant) 
            with 8his tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6031)
  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 6031)
  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-SARS-CoV-2-S-RBD-8his-N501Y
FEATURES             Location/Qualifiers
     source          1..6031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1321..1344
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(1388..1405)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1394..1618
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1664..2092
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1751..1770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1961..1982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2101..2121)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2106..2435
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2286..2421
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2348..2367
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2502..3296
                     /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(2556..2575)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3166..3185
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3470..3591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3507..3526)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3561..3580
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3640..3656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3640..3656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3653..3675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3664..3680
                     /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(3688..3694,3695..3712,3713..3718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3733..3754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3871..3888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4042..4630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4122..4141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4801..5592,5593..5661))
                     /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     5424..5443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5662..5766)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5841..5860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgcca ccatgaagac catcatcgcc ctgagctaca tcttctgcct ggtgttcgcc
      721 actaatcttt gtccgttcgg tgaggttttt aacgcgacaa ggttcgctag tgtatatgct
      781 tggaaccgaa agagaatctc caattgcgta gctgattact ccgttctcta taacagtgcg
      841 tccttttcaa cctttaagtg ttacggcgtt tctccaacga agctgaatga tctctgtttt
      901 acgaacgtgt atgctgactc tttcgttata cggggggacg aagtgagaca gatagcacca
      961 ggtcagactg ggaagatagc ggattacaac tataagttgc ccgatgattt tacggggtgc
     1021 gtaatcgcat ggaactcaaa caacctcgac tccaaagtag gtggtaatta taattacttg
     1081 tatcgcctgt ttcgaaagag caatttgaag ccttttgagc gggatatttc aaccgaaatt
     1141 taccaagcag gcagtacgcc atgtaacgga gtagagggat ttaattgcta ctttcctctt
     1201 caatcttatg gctttcaacc aacatacgga gtggggtatc aaccttatag agtggtagta
     1261 ttgtcctttg agctcctcca cgccccggct acagtttgtg ggcccaaaaa gggatccggt
     1321 caccatcacc accaccatca tcattgataa ctcgagtcta gagggcccgt ttaaacccgc
     1381 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1441 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1501 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1561 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     1621 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     1681 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1741 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1801 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1861 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1921 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1981 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2041 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2101 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2161 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2221 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2281 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2341 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     2401 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     2461 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2521 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2581 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2641 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     2701 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2761 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     2821 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     2881 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     2941 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3001 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3061 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3121 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3181 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3241 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3301 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3361 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3421 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3481 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3541 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3601 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3661 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3721 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3781 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3841 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3901 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3961 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4441 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4681 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4741 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4861 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4921 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4981 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5221 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5461 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5641 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5701 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5761 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     5821 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     5881 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     5941 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     6001 cttcgcgatg tacgggccag atatacgcgt t
//
