LOCUS       sequence-433279-        3939 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes SARS-CoV-2 spike D1118H for pseudovirus production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3939)
  AUTHORS   Roederer AL, Cao Y, Denis KS, Sheehan ML, Li CJ, Lam EC, Gregory DJ,
            Poznansky MC, Iafrate AJ, Canaday DH, Gravenstein S, Garcia-Beltran 
            WF, Balazs AB
  TITLE     Ongoing evolution of SARS-CoV-2 drives escape from mRNA 
            vaccine-induced humoral immunity.
  JOURNAL   medRxiv [Preprint]. 2024 Mar 7:2024.03.05.24303815. doi: 
            10.1101/2024.03.05.24303815.
  PUBMED    38496628
REFERENCE   2  (bases 1 to 3939)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "medRxiv 
            [Preprint]. 2024 Mar 7:2024.03.05.24303815. doi: 
            10.1101/2024.03.05.24303815."
COMMENT     Sequence Label: pTwist-SARS-CoV-2 Spike-deltaC18 D1118H (CAC)
FEATURES             Location/Qualifiers
     source          1..3939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    209..280
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(325..461)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(474..1265,1266..1334))
                     /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     1097..1116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2080..2668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2569..2588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2864..3088
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(3099..3893)
                     /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(3210..3229)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3820..3839
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(3907..3939,1..164))
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(3921..3939,1..117)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
       61 aattagtcag cgatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
      121 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcagatcgc ttaattaagt
      181 ttcgatagcc caaggtaacc aatgagatta gggagggaag aaagtttgat gaactacttc
      241 tgacccctag tggtgtccag aaaagaccat taaaggaatg tcttgatcga aatcgagaga
      301 tcgcctgcag gatagctgac gaccttaagg ataaatttct ggtaaggagg acacgtatgg
      361 aagtgggcaa gttggggaag ccgtatccgt tgctgaatct ggcatatgtg ggagtataag
      421 acgcgcagcg tcgcatcagg catttttttc tgcgccaatg cacctagaat cggttaccaa
      481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
      541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
      601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
      661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatc
      721 aactgttgcc gggaagctag agtaagtagt tcgccagtta atagtttacg caaggttgtt
      781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      901 tccttcggtc ctccaatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1021 ggtgaatact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1141 ggaaagcgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1201 atgtaaccca ctcgcgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1321 tgttgaatac tcatgcgaaa gcctcctgct gatcctgtct cttgatagat gtttgcaaaa
     1381 gctaggactc gattatacgt tgaccgatca gactatcaat atctaggttt aaacagatcg
     1441 gtacgctgcc gtgaccaggc ttgagtacaa tatctgacga ttggacttcc tgggtcccaa
     1501 gctggattcc gactggctga ctgccaagca acgatatgga tcgtcgagac tcgcactgag
     1561 aggatcgttc tcggtacacc aggcgcgccc cgatttcgat attcctcgga tctcggatag
     1621 atcgggattg aggatgtagg ttgaacttct ctcgggtgat cccgatctcg gatattcggg
     1681 attggagtaa tacctggcga taggcgctgc caagcaatag ccaaggcgca aggccggcca
     1741 agagacttgc gatagttcgt tcgagaggta gcctgcaaga tggatcggac gaaaggtttg
     1801 aagattttcc gacgagggcc ttggagagtt cggaaattta aatgcaacta tggatgaacg
     1861 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1921 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1981 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2041 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2101 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2161 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2221 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2281 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2341 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2401 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2461 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2521 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2581 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2641 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2701 ggccttttgc tggccttttg ctcacatgat ttaaatgaaa ctcgtaatcg gagaacgggt
     2761 gcaatagcaa ggaccagatc gaacttgact gacgcaattc tgagcgtagt cgaaacccag
     2821 acaagtcgat ctcgccaagt agctgacttg actggttaat taaccataga gcccaccgca
     2881 tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca
     2941 ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag
     3001 gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca
     3061 acagatggct ggcaactaga aggcacaggg cttacatact agaagaactc gtcaagaagg
     3121 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     3181 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     3241 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     3301 accatgatat tcggcaagca ggcatcgccg tgggtcacga cgagatcctc gccgtcgggc
     3361 atccgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     3421 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     3481 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     3541 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     3601 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     3661 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctggagttca
     3721 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     3781 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     3841 ctctccaccc aagcggcggg agaacctgcg tgcaatccat cttgttcaat catgttggct
     3901 acctcctttg caaaagccta ggcctccaaa aaagcctcc
//
