LOCUS       sequence-409578-        9190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expressing XBB.1.5+K478R SARS-CoV-2 
            Spike, with 19AA cytoplasmic tail truncation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9190)
  TITLE     SARS-CoV-2 PSV spike variants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9190)
  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: 
            "Unpublished"
COMMENT     Sequence Label: XBB.1.5+K478R
FEATURES             Location/Qualifiers
     source          1..9190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4781..4798)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4787..5011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5057..5485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5144..5163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5354..5375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5494..5514)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5499..5828
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5679..5814
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5895..6689
                     /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(5949..5968)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6559..6578
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6863..6984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6900..6919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6954..6973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7033..7049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7033..7049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7046..7068)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7057..7073
                     /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(7081..7087,7088..7105,7106..7111))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7126..7147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7264..7281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7435..8023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7515..7534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8194..8985,8986..9054))
                     /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     8817..8836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9055..9159)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttatgg ccaccatgtt cgtgtttctg gtgctgctgc ctctggtgtc
      961 cagccagtgt gtgaacctga tcaccagaac acagtcgtac accaacagct ttaccagagg
     1021 cgtgtactac cccgacaagg tgttcagatc cagcgtgctg cactctaccc aggacctgtt
     1081 cctgcctttc ttcagcaacg tgacctggtt ccacgccatc cacgtgtccg gcaccaatgg
     1141 caccaagaga ttcgacaacc ccgcgctgcc cttcaacgac ggggtgtact ttgccagcac
     1201 cgagaagtcc aacatcatca gaggctggat cttcggcacc acactggaca gcaagaccca
     1261 gagcctgctg atcgtgaaca acgccaccaa cgtggtcatc aaagtgtgcg agttccagtt
     1321 ctgcaacgac cccttcctgg acgtctacca gaagaacaac aagagctgga tggaaagcga
     1381 gttccgggtg tacagcagcg ccaacaactg caccttcgag tacgtgtccc agcctttcct
     1441 gatggacctg gaaggcaagg agggcaactt caagaacctg cgcgagttcg tgttcaagaa
     1501 catcgacggc tacttcaaga tctacagcaa gcacacccct atcaacctcg agcgggatct
     1561 gcctcagggc ttctctgctc tggaacccct ggtggatctg cccatcggca tcaacatcac
     1621 ccggtttcag acactgctgg ccctgcacag aagctacctg acacctgtcg atagcagcag
     1681 cggatggaca gctggtgccg ccgcttacta tgtgggctac ctgcagccta gaaccttcct
     1741 gctgaagtac aacgagaacg gcaccatcac cgacgccgtg gattgtgccc ttgatcctct
     1801 gagcgagaca aagtgcaccc tgaagtcctt caccgtggaa aagggcatct accagaccag
     1861 caacttccgg gtgcagccca ccgaatccat cgtgcggttc cccaatatca ccaatctgtg
     1921 ccccttccac gaggtgttca atgccaccac attcgcctct gtgtacgcct ggaaccggaa
     1981 gcggatcagc aattgcgtgg ccgactactc cgtgatctac aacttcgccc ccttcttcgc
     2041 cttcaagtgc tacggcgtgt cccctaccaa gctgaacgac ctgtgcttca caaacgtgta
     2101 cgccgacagc ttcgtgatcc ggggaaatga agtgagtcag attgcccctg gacagacagg
     2161 caacatcgcc gactacaact acaagctgcc cgacgacttc accggctgtg tgattgcctg
     2221 gaacagcaac aagctggact ccaaacccag cggcaactac aattacctgt accggctgtt
     2281 ccggaagtcc aagctgaagc ccttcgagcg ggacatctcc accgagatct atcaggccgg
     2341 caaccggcct tgtaacggcg tggcaggccc caactgctac tccccactgc agtcctacgg
     2401 ctttcggccc acatatggcg tgggccatca gccctacaga gtggtggtgc tgagcttcga
     2461 actgctgcat gcccctgcca cagtgtgcgg ccctaagaaa agcaccaatc tcgtgaagaa
     2521 caaatgcgtg aacttcaact tcaacggcct gaccggcaca ggcgtgctga cagagagcaa
     2581 caagaagttc ctgccattcc agcagtttgg ccgggatatc gccgatacca cagacgccgt
     2641 tagagatccc cagacactgg aaatcctgga catcacccct tgcagcttcg gcggagtgtc
     2701 tgtgatcacc cctggcacca acaccagcaa tcaggtggca gtgctgtacc agggcgtgaa
     2761 ctgtaccgaa gtgcccgtgg ccattcacgc cgatcagctg acacctacat ggcgggtgta
     2821 ctccaccggc agcaatgtgt ttcagaccag agccggctgt ctgatcggag ccgagtacgt
     2881 gaacaatagc tacgagtgcg acatccccat cggcgctggc atctgtgcca gctaccagac
     2941 acagacaaag agccacagac gggccagatc tgtggccagc cagagcatca ttgcctacac
     3001 aatgtctctg ggcgccgaga acagcgtggc ctactccaac aactctatcg ctatccccac
     3061 caacttcacc atcagcgtga ccacagagat cctgcctgtg tccatgacca agaccagcgt
     3121 ggactgcacc atgtacatct gcggcgattc caccgagtgc tccaacctgc tgctgcagta
     3181 cggcagcttc tgcacccagc tgaaaagagc cctgacaggg atcgccgtgg aacaggacaa
     3241 gaacacccaa gaggtgttcg cccaagtgaa gcagatctac aagacccctc ctatcaagta
     3301 cttcggcggc ttcaatttca gccagattct gcccgatcct agcaagccca gcaagcggag
     3361 cttcatcgag gacctgctgt tcaacaaagt gacactggcc gacgccggct tcatcaagca
     3421 gtatggcgat tgtctgggcg acattgccgc cagggatctg atttgcgccc agaagtttaa
     3481 cggactgaca gtgctgcctc ctctgctgac cgatgagatg atcgcccagt acacatctgc
     3541 cctgctggcc ggcacaatca caagcggctg gacatttgga gctggcgccg ctctgcagat
     3601 cccctttgct atgcagatgg cctacagatt caacggcatc ggagtgaccc agaatgtgct
     3661 gtacgagaac cagaagctga tcgccaacca gttcaacagc gccatcggca agatccagga
     3721 cagcctgagc agcacagcaa gcgccctggg aaagctgcag gacgtggtca accacaatgc
     3781 ccaggcactg aacaccctgg tcaagcagct gtcctccaag ttcggcgcca tcagctctgt
     3841 gctgaacgat atcctgagca gactggacaa ggtggaggcc gaggtgcaga tcgacagact
     3901 gatcacaggc agactgcaga gcctccagac atacgtgacc cagcagctga tcagagccgc
     3961 cgagattaga gcctctgcca atctggccgc caccaagatg tctgagtgtg tgctgggcca
     4021 gagcaagaga gtggactttt gcggcaaggg ctaccacctg atgagcttcc ctcagtctgc
     4081 ccctcacggc gtggtgtttc tgcacgtgac atacgttccc gctcaagaga agaatttcac
     4141 caccgctcca gccatctgcc acgacggcaa agcccacttt cctagagaag gcgtgttcgt
     4201 gtccaacggc acccattggt tcgtgacaca gcggaacttc tacgagcccc agatcatcac
     4261 caccgacaac accttcgtgt ctggcaactg cgacgtcgtg atcggcattg tgaacaatac
     4321 cgtgtacgac cctctgcagc ccgagctgga cagcttcaaa gaggaactgg acaagtactt
     4381 taagaaccac acaagccccg acgtggacct gggcgatatc agcggaatca atgccagcgt
     4441 cgtgaacatc cagaaagaga tcgaccggct gaacgaggtg gccaagaatc tgaacgagag
     4501 cctgatcgac ctgcaagaac tggggaagta cgagcagtac atcaagtggc cctggtacat
     4561 ctggctgggc tttatcgccg gactgattgc catcgtgatg gtcacaatca tgctgtgttg
     4621 catgaccagc tgctgtagct gcctgaaggg ctgttgtagc tgtggcagct gctgctagtt
     4681 cgacgaggac gattctgagc ccgtgctgaa gggcgtgaaa ctgcactaca catgagcggc
     4741 cgcctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     4801 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4861 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4921 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4981 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     5041 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5101 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5161 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5221 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5281 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5341 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5401 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5461 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     5521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5581 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5641 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5701 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5761 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5821 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5881 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5941 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6001 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6061 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6121 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6181 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6241 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6301 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6361 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6421 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6481 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6541 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6601 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6661 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     6721 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     6781 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6841 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6901 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6961 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     7021 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7081 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7141 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7201 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7261 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7321 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7381 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7441 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7501 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7561 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7621 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7681 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7741 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7801 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7861 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7921 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7981 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8041 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8101 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8161 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8221 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8281 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8341 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8401 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8461 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8521 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8581 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8641 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8701 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8761 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8821 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8881 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8941 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9001 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9061 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9121 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9181 acctgacgtc
//
