LOCUS       sequence-381635-        9197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressing SARS-CoV-2 BA2.75.2 spike protein for pseudovirus 
            production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9197)
  TITLE     SARS-CoV-2 variants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9197)
  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            "Unpublished"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: pcDNA3.3_SARS2_BA.2.75.2
FEATURES             Location/Qualifiers
     source          1..9197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     588..612
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(4541..4560)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4585..4633
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4835..5263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4922..4941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5132..5153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5272..5292)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5277..5606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5457..5592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5519..5538
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5673..6467
                     /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(5727..5746)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6337..6356
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6643..6764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6680..6699)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6734..6753
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6813..6829)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6813..6829)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6826..6848)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6837..6853
                     /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(6861..6867,6868..6885,6886..6891))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6906..6927
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7044..7061)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7215..7803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7295..7314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7974..8765,8766..8834))
                     /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     8597..8616
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8835..8939)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9014..9033)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      601 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      661 gaccgatcca gcctccggac tgagctcctt aaggttaact ctagagaatt cggatccatg
      721 tttgtgttcc tggtgctgct gccactggtg tccagccagt gtgtgaacct gatcaccagg
      781 acccaatcct acaccaactc cttcaccagg ggagtctact accctgacaa ggtgttcagg
      841 tcctctgtgc tgcacagcac ccaggacctg ttcctgccat tcttcagcaa tgtgacctgg
      901 ttccatgcca tccatgtgtc tggcaccaat ggcaccaaga ggtttgacaa ccctgtgctg
      961 ccattcaatg atggagtcta ctttgccagc acagagaaga gcaacatcat caggggctgg
     1021 atttttggca ccaccctgga cagcaagacc cagtccctgc tgattgtgaa caatgccacc
     1081 aatgtggtca ttaaggtgtg tgagttccag ttctgtaatg acccattcct ggacgtctac
     1141 taccacgaga acaacaagtc caggatggag tctgagctca gggtctactc ctctgccaac
     1201 aactgtacct ttgaatatgt gagccaacca ttcctgatgg acttggaggg caagcagggc
     1261 aacttcaaga acctgaggga gtttgtgttc aagaacattg atggctactt caagatttac
     1321 agcaaacaca caccagtcaa cctggggagg gacctgccac agggcttctc tgccttggaa
     1381 ccactggtgg acctgccaat tggcatcaac atcaccaggt tccagaccct gctggctctg
     1441 cacaggtcct acctgacacc tggagactcc agctcttcat ggacagcagg agcagcagcc
     1501 tactatgtgg gctacctcca accaaggacc ttcctgctga aatacaatga gaatggcacc
     1561 atcacagatg ctgtggactg tgccctggac ccactgtctg agaccaagtg taccctgaaa
     1621 tccttcacag tggagaaggg catctaccag accagcaact tcagggtcca accaacagag
     1681 agcattgtga ggtttccaaa catcaccaac ctgtgtccat ttcacgaggt gttcaatgcc
     1741 accacgtttg cctctgtcta tgcctggaac aggaagagga ttagcaactg tgtggctgac
     1801 tactctgtgc tctacaactt tgcccccttc ttcgccttca agtgttatgg agtgagccca
     1861 accaaactga atgacctgtg tttcaccaat gtctatgctg actcctttgt gattagggga
     1921 aatgaggtga gtcagattgc ccctggacaa acaggcaaca ttgctgacta caactacaaa
     1981 ctgcctgatg acttcacagg ctgtgtgatt gcctggaact ccaacaagct ggacagcaag
     2041 gtgtcaggca actacaacta cctctacaga ctgttcagga agagcaagct gaaaccattt
     2101 gagagggaca tcagcacaga gatttaccag gctggcaaca aaccatgtaa tggagtggcg
     2161 ggctccaact gttactttcc actccagtcc tatggcttca gaccaaccta tggagtgggc
     2221 caccaaccat acagggtggt cgtgctgtcc tttgaactgc tccatgcccc tgccacagtg
     2281 tgtggaccaa agaagagcac caacctggtg aagaacaagt gtgtgaactt caacttcaat
     2341 ggactgacag gcacaggagt gctgacagag agcaacaaga agttcctgcc attccaacag
     2401 tttggcaggg acattgctga caccacagat gctgtgaggg acccacagac cttggagatt
     2461 ctggacatca caccatgttc ctttggagga gtgtctgtga ttacacctgg caccaacacc
     2521 agcaaccagg tggctgtgct ctaccagggt gtgaactgta ctgaggtgcc tgtggctatc
     2581 catgctgacc aacttacacc aacctggagg gtctacagca caggcagcaa tgtgttccag
     2641 accagggctg gctgtctgat tggagcagag tatgtgaaca actcctatga gtgtgacatc
     2701 ccaattggag caggcatctg tgcctcctac cagacccaga ccaagagcca taggagggca
     2761 aggtctgtgg caagccagag catcattgcc tacacaatga gtctgggagc agagaactct
     2821 gtggcttaca gcaacaacag cattgccatc ccaaccaact tcaccatctc tgtgaccaca
     2881 gaaattctgc ctgtgagtat gaccaagacc tctgtggact gtacaatgta tatctgtgga
     2941 gacagcacag agtgtagcaa cctgctgctc caatatggct ccttctgtac ccaacttaag
     3001 agggctctga caggcattgc tgtggaacag gacaagaaca cccaggaggt gtttgcccag
     3061 gtgaagcaga tttacaagac acctccaatc aagtactttg gaggcttcaa cttcagccag
     3121 attctgcctg acccaagcaa gccaagcaag aggtccttca ttgaggacct gctgttcaac
     3181 aaggtgaccc tggctgatgc tggcttcatc aagcaatatg gagactgtct gggagacatt
     3241 gctgcaaggg acctgatttg tgcccagaag ttcaatggac tgaccgtgct gcctccactg
     3301 ctgacagatg agatgattgc ccaatacacc tctgccctgc tggctggcac catcacctct
     3361 ggctggacct ttggagcagg agcagccctc caaatcccat ttgctatgca gatggcttac
     3421 aggttcaatg gcattggagt gacccagaat gtgctctatg agaaccagaa actgattgcc
     3481 aaccagttca actctgccat tggcaagatt caggactccc tgtccagcac agcctctgcc
     3541 ctgggcaaac tccaagatgt ggtgaaccat aatgcccagg ctctgaacac cctggtgaag
     3601 caactttcca gcaagtttgg agccatctcc tctgtgctga atgacatcct gagcagactg
     3661 gacaaggtgg aggctgaggt ccagattgac agactgatta caggcagact ccaatccctc
     3721 caaacctatg tgacccaaca acttatcagg gctgctgaga ttagggcatc tgccaacctg
     3781 gctgccacca agatgagtga gtgtgtgctg ggacaaagca agagggtgga cttctgtggc
     3841 aagggctacc acctgatgag ttttccacag tctgcccctc atggagtggt gttcctgcat
     3901 gtgacctatg tgcctgccca ggagaagaac ttcaccacag cccctgccat ctgccatgat
     3961 ggcaaggctc actttccaag ggagggagtg tttgtgagca atggcaccca ctggtttgtg
     4021 acccagagga acttctatga accacagatt atcaccacag acaacacctt tgtgtctggc
     4081 aactgtgatg tggtgattgg cattgtgaac aacacagtct atgacccact ccaacctgaa
     4141 ctggactcct tcaaggagga actggacaaa tacttcaaga accacaccag ccctgatgtg
     4201 gacctgggag acatctctgg catcaatgcc tctgtggtga acatccagaa ggagattgac
     4261 agactgaatg aggtggctaa gaacctgaat gagtccctga ttaacctcca agaactgggc
     4321 aaatatgaac aatacatcaa gtggccatgg tacatctggc tgggcttcat tgctggactg
     4381 attgccattg tgatggtgac cataatgctg tgttgtatga cctcctgttg ttcctgtctg
     4441 aaaggctgtt gttcctgtgg ctcctgttgt aagtgactcg agccatggaa gcttgatatc
     4501 taactgactg aaccggtggt accaaacggg ggaggctaac tgaaacacgg aaggagacaa
     4561 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     4621 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4681 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4741 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     4801 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4861 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4921 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4981 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5041 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5101 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5161 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5221 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5281 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5341 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5401 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5461 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5521 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5581 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5641 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5701 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5761 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5821 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5881 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5941 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6001 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6061 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6121 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6181 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6241 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6301 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6361 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6421 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6481 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6541 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6601 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6661 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6721 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6781 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6841 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6901 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6961 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7021 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7081 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7141 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7201 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7261 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7321 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7381 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7441 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7501 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7561 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7621 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7681 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7741 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7801 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7861 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7921 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7981 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8041 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8101 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8161 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8221 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8281 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8341 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8401 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8461 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8521 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8581 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8641 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8701 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8761 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8821 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8881 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8941 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     9001 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     9061 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     9121 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     9181 atgtacgggc cagatat
//
