LOCUS       sequence-306267-        7481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 surface glycoprotein 
            (Spike) for VSV pseudotyping.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7481)
  TITLE     Truncated SARS-CoV-2 Spike protein constructs for making VSV pseudo 
            virus
REFERENCE   2  (bases 1 to 7481)
  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"
COMMENT     Sequence Label: pGBW-m4137384
FEATURES             Location/Qualifiers
     source          1..7481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        136..515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        516..719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     670..690
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(4634..4653)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4699..4754
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4753..4772)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5115..5131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5115..5131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5128..5150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5139..5155
                     /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(5163..5169,5170..5187,5188..5193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5208..5229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5288..5483
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5334..5469
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5489..5623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5526..5545)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5580..5599
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5691..5708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5862..6450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5942..5961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6621..6689,6690..7481)
                     /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(6839..6858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gggtcgacat tgattattga ctagttatta atagtaatca attacggggt
      181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      301 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      481 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      541 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      601 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      661 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
      721 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
      781 agcggccgcc accatgttcg tctttcttgt gttactcccc ctggtttcgt cccagtgcgt
      841 gaacttgact acgcgaaccc aactgccgcc agcatataca aatagtttca cccgtggagt
      901 atactaccct gataaagtgt ttagatcaag cgtactacac tctactcagg acctgtttct
      961 ccccttcttc tccaatgtca catggtttca tgcgatccac gttagcggca ccaacgggac
     1021 aaagcgcttt gacaaccctg tgctgccatt caatgatggt gtgtattttg cttctaccga
     1081 gaagagcaac ataattcggg gctggatctt cggaactacc ctcgacagta aaactcagtc
     1141 attgctgatt gtgaataacg ccacaaatgt cgttatcaag gtctgtgaat ttcaattctg
     1201 caacgatcca ttccttgggg tgtattacca taaaaataat aagtcctgga tggagagcga
     1261 atttagggta tactcgtctg ccaacaattg tactttcgag tatgtgtctc agcccttctt
     1321 aatggatctg gaaggtaaac agggcaactt taagaacctc cgcgagtttg tattcaaaaa
     1381 tatagacgga tactttaaga tttattccaa gcacacacca atcaatttgg tgcgggatct
     1441 cccgcaaggg ttcagcgctc tggaacctct agtggacctc cccatcggca ttaacatcac
     1501 aagattccag acgctgttag ccctgcatag atcatacctt actcccggtg acagtagttc
     1561 tgggtggacc gcaggcgcgg ctgcatatta cgtgggatat ttgcagcctc ggacgtttct
     1621 gctgaaatac aacgagaacg gcaccattac agatgccgtg gattgcgccc ttgaccccct
     1681 aagcgaaact aagtgtacgc ttaaaagctt caccgtcgag aagggaattt accagacttc
     1741 caactttagg gtccagccga cagaatcaat agttaggttc ccaaatatca ccaatttgtg
     1801 cccttttggg gaagtcttca atgctacacg atttgcttcg gtttatgcct ggaacaggaa
     1861 gcgtatctca aattgcgtcg cggactacag cgtgctctat aatagtgcaa gtttctcgac
     1921 ctttaaatgt tatggcgtgt ccccgaccaa actaaatgac ctctgtttta caaacgttta
     1981 cgccgattct tttgtgattc gcggtgatga ggtacgccaa atcgcaccgg gccagactgg
     2041 caagattgcc gattacaact acaagttgcc tgacgacttc accggatgtg ttatagcctg
     2101 gaatagcaat aatctggatt ctaaggtcgg aggtaactat aactacctgt atagactgtt
     2161 ccgtaagtca aacctgaaac cctttgagcg ggacatctcc acagagattt accaagcagg
     2221 gtctaccccc tgcaatgggg ttgaaggatt caattgctac tttccattac agagctacgg
     2281 gttccagcct acgaacgggg ttggttacca gccttatagg gtagtggtgc tgtcgttcga
     2341 actgttgcac gctccagcta cagtgtgcgg gcctaaaaaa tcaactaacc tggttaaaaa
     2401 taagtgtgtt aattttaact tcaatggctt aaccggaact ggagtgctca cggagtcaaa
     2461 caagaaattt ctaccgtttc agcaattcgg gagggatatc gcagacacta cggacgcggt
     2521 cagagatcca cagacccttg agatccttga cataacaccc tgctctttcg gaggtgtctc
     2581 cgtaattaca ccaggcacta atacctctaa tcaggtggcc gtgctttatc aggatgttaa
     2641 ctgtacagag gtcccggtgg ctatccatgc cgaccaattg acacctacgt ggcgcgtcta
     2701 ttctaccggc tccaacgtct tccagacccg ggctggatgc ttgataggcg cagaacacgt
     2761 aaacaactct tacgaatgtg atattcccat tggagccggg atctgcgcaa gctaccagac
     2821 ccaaacaaac agtcccagac gggctcgtag cgtggcctct cagagcataa tagcctatac
     2881 catgtccctt ggcgcggaga atagtgtggc atactcaaat aatagcatcg ctatccccac
     2941 taactttact atttccgtga cgactgagat tctgccagtg tctatgacta agacatccgt
     3001 ggattgcaca atgtatattt gtggggattc cacagagtgt agcaacttgt tgctgcaata
     3061 tggatcattc tgtacgcagc tgaatcgcgc cctcaccggc atcgctgtgg aacaggacaa
     3121 gaacactcaa gaggtctttg ctcaggtgaa gcaaatctac aagaccccac ctataaaaga
     3181 cttcgggggt tttaacttct cgcagattct ccccgacccc agcaagcctt caaaaaggag
     3241 ctttattgag gacctgttgt tcaacaaggt tactttagcc gatgccggtt ttatcaaaca
     3301 gtacggcgac tgtcttggcg atatagcagc ccgggatctg atttgcgcac aaaaattcaa
     3361 tggccttacc gtactgccac cccttctcac cgacgaaatg atcgcgcagt acacctccgc
     3421 actgctggct gggaccatca cttcgggatg gacctttggg gctggtgctg ctttgcaaat
     3481 tcccttcgcc atgcagatgg cctatcgatt caatggcata ggggtaacac agaacgtatt
     3541 gtatgaaaat caaaagctaa tcgctaacca gtttaattca gccattggta agatccagga
     3601 ttcactctcc agtacagcct ctgccctggg aaagctccag gacgtggtca accagaacgc
     3661 gcaggcattg aatacactcg tcaaacagct gagtagcaat ttcggcgcaa tcagtagtgt
     3721 cttaaacgat atactgagta ggcttgataa agtagaggcc gaggtccaga ttgacagact
     3781 aatcacgggg cgtcttcaga gtttacagac ctatgtgacc caacagctga tccgagcagc
     3841 ggaaattcga gcaagcgcga acctggccgc tactaaaatg agtgagtgcg tgctgggcca
     3901 aagtaaacga gtcgactttt gcggcaaagg gtatcatctc atgtctttcc ctcagtccgc
     3961 ccctcacggt gttgttttcc tccacgtgac atacgttccc gcgcaggaaa agaattttac
     4021 tacggcaccc gctatttgcc atgacggaaa ggctcatttt ccacgggagg gagtgttcgt
     4081 ttcaaacggg acacactggt ttgtaactca gcggaacttt tatgagcctc aaatcatcac
     4141 gacagacaat accttcgtga gtggcaactg cgatgtagtc ataggcattg tgaataacac
     4201 tgtttatgat ccacttcagc cggaactcga cagcttcaaa gaggaattgg ataagtactt
     4261 taaaaatcat accagtccag acgtagatct cggcgatata tcgggcatta acgccagtgt
     4321 ggtcaatata caaaaggaga ttgacagact gaatgaagta gctaagaacc tcaacgaatc
     4381 tctgattgac ctgcaagagc tagggaagta cgagcagtat atcaaatggc catggtacat
     4441 atggctgggt tttatcgcgg gtcttattgc catagtcatg gtgacgatta tgttgtgctg
     4501 tatgacctcc tgctgttctt gtttgaaagg ctgctgctcc tgtggtagct gttgcaaatt
     4561 cgatgaagat gactctgaac ccgtgttgaa aggcgttaag ctacactaca cataatccgg
     4621 actcagatct cgatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     4681 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4741 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     4801 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     4861 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     4921 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     4981 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     5041 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc
     5101 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5161 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5221 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5281 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5341 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5401 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5461 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
     5521 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5581 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
     5641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6301 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6601 gagtaaactt ggtctgacag atgagtattc aacatttccg tgtcgccctt attccctttt
     6661 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6721 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6781 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6841 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6901 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6961 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     7021 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     7081 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7141 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     7201 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7261 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7321 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7381 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7441 agatcgctga gataggtgcc tcactgatta agcattggta a
//
