LOCUS       sequence-297245-        9018 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 S (surface 
            glycoprotein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9018)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9018)
  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"
COMMENT     Sequence Label: pGBW-m4164341
FEATURES             Location/Qualifiers
     source          1..9018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3863..3984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3874..3893)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3928..3947
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4099..4145
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4175..4311
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4356..4427
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4653..5241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4733..4752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5987..6781
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5987..6781
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6041..6060)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6651..6670
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6794..6930
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7075..7146
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7151..7197)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7215..7263
                     /note="synthetic polyadenylation signal"
     enhancer        7346..7649
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7652..7927
                     /label=chicken beta-actin promoter
     intron          7928..8938
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     8862..8884
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 accatgtttg ttttcttggt tcttttgcca ctggtaagca gtcagtgcgt caacctgacc
       61 acccggacac aactcccacc cgcatacacg aacagtttca ctaggggggt ttactaccct
      121 gacaaagttt ttagatctag tgttttgcat tccacgcaag atctgttcct tccgttcttt
      181 tcaaatgtca cgtggttcca tgctattcac gtcagcggca ctaatggaac gaaacgattc
      241 gataaccccg tcttgccctt caacgatggc gtatatttcg cgtcaacaga gaaaagtaat
      301 ataattagag gctggatctt tggtacgact ctcgacagta agacgcagtc acttctgatt
      361 gtaaacaatg caactaatgt agtcatcaag gtgtgtgagt tccagttctg caatgatcct
      421 ttcttgggag tgtattatca taagaataac aaatcctgga tggagtctga gttccgggta
      481 tattcttcag caaacaattg cacgtttgaa tatgtaagcc aacctttcct tatggacctg
      541 gaagggaagc aaggtaattt caagaatctt agggaatttg tgtttaaaaa tatagatggc
      601 tacttcaaga tctactcaaa gcataccccg attaacctcg tcagagacct gccccaaggg
      661 ttcagtgcgc ttgaacccct ggtagatttg ccaatcggca taaatataac caggttccag
      721 acactgctcg ccctccaccg gagctacctt actccaggag attcatctag cgggtggaca
      781 gctggagcgg cagcttacta cgtcggttac cttcaaccga ggacattctt gcttaaatac
      841 aatgagaatg gcacgataac tgacgccgta gattgcgcgc ttgatccgct gtcagaaacg
      901 aagtgtacac ttaaaagttt tacggtcgaa aagggaatct accaaacatc aaattttagg
      961 gtacaaccta cggaatctat cgtacgcttc ccgaatatca ctaacttgtg tccgttcggc
     1021 gaggttttta atgcgaccag gtttgcttcc gtgtacgcct ggaacaggaa acggatctcc
     1081 aattgtgtcg ccgattactc cgtcttgtat aattcagcat ctttcagcac gtttaaatgt
     1141 tacggagttt cccccacaaa attgaatgac ctttgcttta cgaacgtcta cgcggattca
     1201 tttgtaatcc ggggggacga agttaggcaa attgcgccag ggcagactgg caagatagct
     1261 gactataatt ataaattgcc ggatgacttt acgggctgtg tgattgcttg gaactcaaat
     1321 aatctggact caaaggtagg gggaaattat aactaccttt acaggctgtt ccggaagagt
     1381 aatctgaagc cattcgaaag agatataagt acagagatct accaagctgg aagcaccccc
     1441 tgcaatggtg ttgaaggatt caattgttat ttcccattgc aatcctatgg ttttcaaccg
     1501 acgaatgggg tgggatacca accatatcga gttgtggttc tcagtttcga gttgcttcat
     1561 gctcctgcga cagtatgtgg accaaaaaaa tctactaatc tggtgaagaa taaatgcgtc
     1621 aattttaatt ttaatggtct gacaggtacc ggagtgctta cggaatctaa caagaagttt
     1681 cttccttttc aacagttcgg gagggacata gccgacacaa ctgacgcggt aagagatccg
     1741 cagacgttgg aaattctcga tataactcca tgctcatttg gaggagtgag tgttatcact
     1801 cctggtacga acaccagtaa tcaggttgca gttctttatc aggacgtgaa ttgtacagag
     1861 gtcccggttg cgatacacgc ggaccagctg acgcccacgt ggcgggttta ctcaactggg
     1921 tcaaatgtgt ttcaaaccag agccggctgc ctcataggag ccgagcatgt gaacaacagt
     1981 tatgaatgtg acatacctat cggggcgggg atctgcgctt catatcaaac gcaaaccaat
     2041 tccccgcgaa gagcgcgaag tgttgcaagt caaagtatta tagcgtatac gatgagcttg
     2101 ggtgctgaga actccgtggc atattcaaat aattccatcg caattcccac gaattttacg
     2161 atttctgtga cgacggagat attgccggtt tcaatgacta agacgagtgt tgactgtacg
     2221 atgtacatct gcggcgactc aacggagtgt tccaacctgc tcctgcagta cggctcattc
     2281 tgcacccaac tgaatcgggc acttacgggt attgctgttg aacaagacaa aaatacgcaa
     2341 gaggtgttcg cccaggtcaa gcagatctac aagacaccac cgattaagga ctttgggggg
     2401 tttaattttt ctcagatact tcccgatccc agtaaaccta gtaaaaggtc atttatagag
     2461 gacctcttgt tcaacaaggt aacgctcgct gatgcaggat tcatcaagca gtatggcgat
     2521 tgtcttgggg acattgcggc gcgcgacctg atttgtgccc agaagttcaa cggactgacg
     2581 gtacttccgc ctctcctgac cgacgaaatg atcgcacaat acacttctgc attgcttgct
     2641 ggcaccataa cctcaggttg gaccttcggt gccggggcgg ccttgcaaat cccctttgct
     2701 atgcaaatgg catatcgatt caatgggata ggagttactc aaaatgttct gtatgaaaac
     2761 caaaagctga tagccaatca atttaatagt gctattggga aaattcagga cagcctgagt
     2821 agtacggctt ccgctctcgg caaactccaa gatgtcgtta accaaaatgc acaagctctc
     2881 aatacacttg ttaagcaact gagctccaac ttcggcgcta tatcatcagt tcttaacgat
     2941 attctttccc gactcgacaa ggtcgaagcc gaagtccaaa tagacaggct gataaccggg
     3001 aggctgcaaa gtctccaaac gtatgttacc caacaactca tacgagcggc cgagattagg
     3061 gcgtcagcca acctcgccgc tactaaaatg tctgaatgtg tactcgggca gtccaagcgc
     3121 gtggactttt gtggcaaggg ttatcatctg atgagtttcc cgcaatcagc gcctcacgga
     3181 gtggtcttcc tccacgtgac gtatgtccct gcgcaagaaa aaaactttac gacagcccct
     3241 gccatttgtc atgacgggaa ggcgcacttt ccacgcgaag gcgtttttgt atccaacggc
     3301 acccactggt ttgtcacaca gcgaaatttt tatgagcctc agatcataac tactgacaat
     3361 actttcgtct cagggaactg cgatgtagtc atagggatcg taaacaatac ggtatacgat
     3421 cccctccagc ctgaactgga ctcatttaaa gaggagctgg acaagtattt caaaaatcac
     3481 acatcaccgg acgtggacct cggagatatt agtggtatta acgcaagcgt agtaaatatt
     3541 caaaaggaaa tcgaccgctt gaatgaagtc gcgaaaaacc tgaacgagtc tcttattgat
     3601 cttcaagaac tcggtaagta tgaacaatac atcaagtggc cttggtacat atggcttgga
     3661 ttcattgcgg ggttgatcgc aatcgtaatg gttaccatta tgttgtgttg catgacaagc
     3721 tgttgcagct gcctcaaagg ttgttgttct tgcggtagtt gctgcaagtt cgacgaagat
     3781 gattctgaac ctgttcttaa aggcgtaaaa ttgcactaca cgtaagactg agatacagcg
     3841 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3901 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3961 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4021 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     4081 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     4141 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     4201 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     4261 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     4321 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     4381 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     4441 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     4501 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     4561 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     4621 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4741 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4801 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4861 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4921 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4981 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5041 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5101 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5341 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5461 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     5521 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     5581 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     5641 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     5701 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct
     5761 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     5821 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     5881 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     5941 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga ttgaacaaga
     6001 tggattgcac gcaggttctc cggcggcttg ggtggagagg ctattcggct atgactgggc
     6061 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgtcc
     6121 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     6181 gcggctatcg tggctggcga cgacgggcgt tccttgcgcg gctgtgctcg acgttgtcac
     6241 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     6301 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     6361 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     6421 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     6481 cgcgccagcc gaactgttcg ccaggctcaa ggcgtctatg cccgacggcg aggatctcgt
     6541 cgtgacccac ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     6601 attcatcgac tgtggccgtc tgggtgtggc ggaccgctat caggacatag cgttggctac
     6661 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttccttg tgctttacgg
     6721 tatcgccgcg cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6781 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     6841 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     6901 gtgtcctcct taccagaaat ttatccttaa ggtcgtttaa actcgactct ggctctatcg
     6961 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     7021 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     7081 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     7141 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     7201 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     7261 attatctaac tgctgatcga gtgtagccag atctatagta atcaattacg gggtcattag
     7321 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     7381 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     7441 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg
     7501 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     7561 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     7621 tctacgtatt agtcatcgct attaccatgc gtcgaggtga gccccacgtt ctgcttcact
     7681 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     7741 tatgcagcga tgggggcggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga
     7801 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
     7861 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
     7921 gcgggcggga gtcgctgcgt tgccttcgcc ccgtgccccg ctccgcgccg cctcgcgccg
     7981 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     8041 tccctccggg ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc
     8101 gtgaaagcct taaagggctc cgggagggcc tttgtgcggg ggggagcggc tcggggggtg
     8161 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggccc gcgctgcccg gcggctgtga
     8221 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcgtg tgcgcgaggg gagcgcgggc
     8281 cgggggcggt gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt
     8341 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct
     8401 ggcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtgcgg
     8461 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     8521 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc cccggagcgc
     8581 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     8641 ggcgcaggga cttcctttgt cccaaatctg gcggagccga aatctgggag gcgccgccgc
     8701 accccctcta gcgggcgcgg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     8761 agggccttcg tgcgtcgccg cgccgccgtc cccttctcca tctccagcct cggggctgcc
     8821 gcagggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     8881 gaccggcggc tttagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     8941 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagatc tttgtcgatc
     9001 ctaccatcca ctcgacac
//
