LOCUS       sequence-404784-        8307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for SARS-CoV-2 Delta variant spike.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8307)
  AUTHORS   Dadonaite B, Crawford KHD, Radford CE, Farrell AG, Yu TC, Hannon WW,
            Zhou P, Andrabi R, Burton DR, Liu L, Ho DD, Chu HY, Neher RA, Bloom 
            JD
  TITLE     A pseudovirus system enables deep mutational scanning of the full 
            SARS-CoV-2 spike.
  JOURNAL   Cell. 2023 Mar 16;186(6):1263-1278.e20. doi: 
            10.1016/j.cell.2023.02.001. Epub 2023 Feb 13.
  PUBMED    36868218
REFERENCE   2  (bases 1 to 8307)
  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"; doi: 
            "10.1016/j.cell.2023.02.001"; journalName: "Cell"; date: 
            "2023-03-16- 16"; volume: "186"; issue: "6"; pages: "1263-1278.e20"
COMMENT     Sequence Label: HDM_Spikedelta21_sinobiological_B.1.617.2
FEATURES             Location/Qualifiers
     source          1..8307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        462..665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     616..636
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     662..686
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          779..1254
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     CDS             complement(2622..2639)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(5077..5093)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5078..5094)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5187..5581
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(5915..5933)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6001..6105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6106..6174,6175..6966)
                     /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(6324..6343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7137..7725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7626..7645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcttggccc attgcatacg ttgtatccat atcataatat gtacatttat attggctcat
       61 gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
      121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      661 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
      721 atagaagaca ccgggaccga tccagcctcc cctcgaagct gatcctgaga acttcagggt
      781 gagtctatgg gacccttgat gttttctttc cccttctttt ctatggttaa gttcatgtca
      841 taggaagggg agaagtaaca gggtacacat attgaccaaa tcagggtaat tttgcatttg
      901 taattttaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattccg ccaccatgtt
     1321 tgtgttcctg gtgctgctgc cactggtgtc cagccagtgt gtgaacctga gaaccaggac
     1381 ccaacttcct cctgcctaca ccaactcctt caccagggga gtctactacc ctgacaaggt
     1441 gttcaggtcc tctgtgctgc acagcaccca ggacctgttc ctgccattct tcagcaatgt
     1501 gacctggttc catgccatcc atgtgtctgg caccaatggc accaagaggt ttgacaaccc
     1561 tgtgctgcca ttcaatgatg gagtctactt tgccagcaca gagaagagca acatcatcag
     1621 gggctggatt tttggcacca ccctggacag caagacccag tccctgctga ttgtgaacaa
     1681 tgccaccaat gtggtgatta aggtgtgtga gttccagttc tgtaatgacc cattcctgga
     1741 cgtctactac cacaagaaca acaagtcctg gatggagtct ggagtctact cctctgccaa
     1801 caactgtacc tttgaatatg tgagccaacc attcctgatg gacttggagg gcaagcaggg
     1861 caacttcaag aacctgaggg agtttgtgtt caagaacatt gatggctact tcaagattta
     1921 cagcaaacac acaccaatca acctggtgag ggacctgcca cagggcttct ctgccttgga
     1981 accactggtg gacctgccaa ttggcatcaa catcaccagg ttccagaccc tgctggctct
     2041 gcacaggtcc tacctgacac ctggagactc ctcctctggc tggacagcag gagcagcagc
     2101 ctactatgtg ggctacctcc aaccaaggac cttcctgctg aaatacaatg agaatggcac
     2161 catcacagat gctgtggact gtgccctgga cccactgtct gagaccaagt gtaccctgaa
     2221 atccttcaca gtggagaagg gcatctacca gaccagcaac ttcagggtcc aaccaacaga
     2281 gagcattgtg aggtttccaa acatcaccaa cctgtgtcca tttggagagg tgttcaatgc
     2341 caccaggttt gcctctgtct atgcctggaa caggaagagg attagcaact gtgtggctga
     2401 ctactctgtg ctctacaact ctgcctcctt cagcaccttc aagtgttatg gagtgagccc
     2461 aaccaaactg aatgacctgt gtttcaccaa tgtctatgct gactcctttg tgattagggg
     2521 agatgaggtg agacagattg cccctggaca aacaggcaag attgctgact acaactacaa
     2581 actgcctgat gacttcacag gctgtgtgat tgcctggaac agcaacaacc tggacagcaa
     2641 ggtgggaggc aactacaact acagatacag actgttcagg aagagcaacc tgaaaccatt
     2701 tgagagggac atcagcacag agatttacca ggctggcagc aagccatgta atggagtgga
     2761 gggcttcaac tgttactttc cactccaatc ctatggcttc caaccaacca atggagtggg
     2821 ctaccaacca tacagggtgg tggtgctgtc ctttgaactg ctccatgccc ctgccacagt
     2881 gtgtggacca aagaagagca ccaacctggt gaagaacaag tgtgtgaact tcaacttcaa
     2941 tggactgaca ggcacaggag tgctgacaga gagcaacaag aagttcctgc cattccaaca
     3001 gtttggcagg gacattgctg acaccacaga tgctgtgagg gacccacaga ccttggagat
     3061 tctggacatc acaccatgtt cctttggagg agtgtctgtg attacacctg gcaccaacac
     3121 cagcaaccag gtggctgtgc tctaccaggg tgtgaactgt actgaggtgc ctgtggctat
     3181 ccatgctgac caacttacac caacctggag ggtctacagc acaggcagca atgtgttcca
     3241 gaccagggct ggctgtctga ttggagcaga gcatgtgaac aactcctatg agtgtgacat
     3301 cccaattgga gcaggcatct gtgcctccta ccagacccag accaacagca gaaggagggc
     3361 aaggtctgtg gcaagccaga gcatcattgc ctacacaatg agtctgggag cagagaactc
     3421 tgtggcttac agcaacaaca gcattgccat cccaaccaac ttcaccatct ctgtgaccac
     3481 agagattctg cctgtgagta tgaccaagac ctctgtggac tgtacaatgt atatctgtgg
     3541 agacagcaca gagtgtagca acctgctgct ccaatatggc tccttctgta cccaacttaa
     3601 cagggctctg acaggcattg ctgtggaaca ggacaagaac acccaggagg tgtttgccca
     3661 ggtgaagcag atttacaaga cacctccaat caaggacttt ggaggcttca acttcagcca
     3721 gattctgcct gacccaagca agccaagcaa gaggtccttc attgaggacc tgctgttcaa
     3781 caaggtgacc ctggctgatg ctggcttcat caagcaatat ggagactgtc tgggagacat
     3841 tgctgccagg gacctgattt gtgcccagaa gttcaatgga ctgacagtgc tgcctccact
     3901 gctgacagat gagatgattg cccaatacac ctctgccctg ctggctggca ccatcacctc
     3961 tggctggacc tttggagcag gagcagccct ccaaatccca tttgctatgc agatggctta
     4021 caggttcaat ggcattggag tgacccagaa tgtgctctat gagaaccaga aactgattgc
     4081 caaccagttc aactctgcca ttggcaagat tcaggactcc ctgtccagca cagcctctgc
     4141 cctgggcaaa ctccaaaacg tggtgaacca gaatgcccag gctctgaaca ccctggtgaa
     4201 gcaactttcc agcaactttg gagccatctc ctctgtgctg aatgacatcc tgagcagact
     4261 ggacaaggtg gaggctgagg tccagattga cagactgatt acaggcagac tccaatccct
     4321 ccaaacctat gtgacccaac aacttatcag ggctgctgag attagggcat ctgccaacct
     4381 ggctgccacc aagatgagtg agtgtgtgct gggacaaagc aagagggtgg acttctgtgg
     4441 caagggctac cacctgatga gttttccaca gtctgcccct catggagtgg tgttcctgca
     4501 tgtgacctat gtgcctgccc aggagaagaa cttcaccaca gcccctgcca tctgccatga
     4561 tggcaaggct cactttccaa gggagggagt gtttgtgagc aatggcaccc actggtttgt
     4621 gacccagagg aacttctatg aaccacagat tatcaccaca gacaacacct ttgtgtctgg
     4681 caactgtgat gtggtgattg gcattgtgaa caacacagtc tatgacccac tccaacctga
     4741 actggactcc ttcaaggagg aactggacaa atacttcaag aaccacacca gccctgatgt
     4801 ggacctggga gacatctctg gcatcaatgc ctctgtggtg aacatccaga aggagattga
     4861 cagactgaat gaggtggcta agaacctgaa tgagtccctg attgacctcc aagaactggg
     4921 caaatatgaa caatacatca agtggccatg gtacatctgg ctgggcttca ttgctggact
     4981 gattgccatt gtgatggtga ccataatgct gtgttgtatg acctcctgtt gttcctgtct
     5041 gaaaggctgt tgttcctgtg gctcctgaaa gcttatcgat accgtcgacc tcgagggccc
     5101 agatctaatt caccccacca gtgcaggctg cctatcagaa agtggtggct ggtgtggcta
     5161 atgccctggc ccacaagtat cactaagctc gctttcttgc tgtccaattt ctattaaagg
     5221 ttcctttgtt ccctaagtcc aactactaaa ctgggggata ttatgaaggg ccttgagcat
     5281 ctggattctg cctaataaaa aacatttatt ttcattgcaa tgatgtattt aaattatttc
     5341 tgaatatttt actaaaaagg gaatgtggga ggtcagtgca tttaaaacat aaagaaatga
     5401 agagctagtt caaaccttgg gaaaatacac tatatcttaa actccatgaa agaaggtgag
     5461 gctgcaaaca gctaatgcac attggcaaca gcccctgatg cctatgcctt attcatccct
     5521 cagaaaagga ttcaagtaga ggcttgattt ggaggttaaa gttttgctat gctgtatttt
     5581 acattactta ttgttttagc tgtcctcatg aatgtctttt cactacccat ttgcttatcc
     5641 tgcatctctc agccttgact ccactcagtt ctcttgctta gagataccac ctttcccctg
     5701 aagtgttcct tccatgtttt acggcgagat ggtttctcct cgcctggcca ctcagcctta
     5761 gttgtctctg ttgtcttata gaggtctact tgaagaagga aaaacagggg gcatggtttg
     5821 actgtcctgt gagcccttct tccctgcctc ccccactcac agtgacccgg aatccctcga
     5881 catggcagtc tagatcattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     5941 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     6001 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6061 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgga
     7741 gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt
     7801 ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa
     7861 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg
     7921 cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg
     7981 ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa
     8041 gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc
     8101 tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc
     8161 ataatgggga aggccatcca gcctcgcgtc ggggagcttt ttgcaaaagc ctaggcctcc
     8221 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     8281 ataaataaaa aaaattagtc agccatg
//
