LOCUS       sequence-417568-        8609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses IDE fused with 2Strep in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8609)
  AUTHORS   Li J, Gui Q, Liang FX, Sall J, Zhang Q, Duan Y, Dhabaria A, Askenazi
            M, Ueberheide B, Stapleford KA, Pagano M
  TITLE     The REEP5/TRAM1 complex binds SARS-CoV-2 NSP3 and promotes virus 
            replication.
  JOURNAL   J Virol. 2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23.
  PUBMED    37768083
REFERENCE   2  (bases 1 to 8609)
  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: "J Virol. 
            2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23."
COMMENT     Sequence Label: IDE-2Strep
FEATURES             Location/Qualifiers
     source          1..8609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..72)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        244..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        624..827
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     778..798
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     872..891
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        872..890
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3988..3997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3992..4001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4009..4032
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4069..4092
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4117..4140
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(4171..4189)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4172..4189)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4209..4226)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4215..4439
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4485..4913
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4572..4591)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4782..4803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4922..4942)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4927..5256
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5107..5242
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5169..5188
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5323..6117
                     /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(5377..5396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5987..6006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6291..6412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6328..6347)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6382..6401
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6461..6477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6461..6477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6474..6496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6485..6501
                     /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(6509..6515,6516..6533,6534..6539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6554..6575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6692..6709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6863..7451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6943..6962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7622..8413,8414..8482))
                     /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     8245..8264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8483..8587)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
       61 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
      121 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
      181 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
      241 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      661 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      721 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      781 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      841 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      901 cttggtaccg agctcggatc catgcggtac cggctagcgt ggcttctgca ccccgcactg
      961 cccagcacct tccgctcagt cctcggcgcc cgcctgccgc ctccggagcg cctgtgtggt
     1021 ttccaaaaaa agacttacag caaaatgaat aatccagcca tcaagagaat aggaaatcac
     1081 attaccaagt ctcctgaaga caagcgagaa tatcgagggc tagagctggc caatggtatc
     1141 aaagtacttc ttatcagtga tcccaccacg gataagtcat cagcagcact tgatgtgcac
     1201 ataggttcat tgtcggatcc tccaaatatt gctggcttaa gtcatttttg tgaacatatg
     1261 ctttttttgg gaacaaagaa ataccctaaa gaaaatgaat acagccagtt tctcagtgag
     1321 catgcaggaa gttcaaatgc ctttactagt ggagagcata ccaattacta ttttgatgtt
     1381 tctcatgaac acctagaagg tgccctagac aggtttgcac agttttttct gtgccccttg
     1441 ttcgatgaaa gttgcaaaga cagagaggtg aatgcagttg attcagaaca tgagaagaat
     1501 gtgatgaatg atgcctggag actctttcaa ttggaaaaag ctacagggaa tcctaaacac
     1561 cccttcagta aatttgggac aggtaacaaa tatactctgg agactagacc aaaccaagaa
     1621 ggcattgatg taagacaaga gctactgaaa ttccattctg cttactattc atccaactta
     1681 atggctgttt gtgttttagg tcgagaatct ttagatgact tgactaatct ggtggtaaag
     1741 ttattttctg aagtagagaa caaaaatgtt ccattgccag aatttcctga acaccctttc
     1801 caagaagaac atcttaaaca actttacaaa atagtaccca ttaaagatat taggaatctc
     1861 tatgtgacat ttcccatacc tgaccttcag aaatactaca aatcaaatcc tggtcattat
     1921 cttggtcatc tcattgggca tgaaggtcct ggaagtctgt tatcagaact taagtcaaag
     1981 ggctgggtta atactcttgt tggtgggcag aaggaaggag cccgaggttt tatgtttttt
     2041 atcattaatg tggacttgac cgaggaagga ttattacatg ttgaagatat aattttgcac
     2101 atgtttcaat acattcagaa gttacgtgca gaaggacctc aagaatgggt tttccaagag
     2161 tgcaaggact tgaatgctgt tgcttttagg tttaaagaca aagagaggcc acggggctat
     2221 acatctaaga ttgcaggaat attgcattat tatcccctag aagaggtgct cacagcggaa
     2281 tatttactgg aagaatttag acctgactta atagagatgg ttctcgataa actcagacca
     2341 gaaaatgtcc gggttgccat agtttctaaa tcttttgaag gaaaaactga tcgcacagaa
     2401 gagtggtatg gaacccagta caaacaagaa gctataccgg atgaagtcat caagaaatgg
     2461 caaaatgctg acctgaatgg gaaatttaaa cttcctacaa agaatgaatt tattcctacg
     2521 aattttgaga ttttaccgtt agaaaaagag gcgacaccat accctgctct tattaaggat
     2581 acagctatga gcaaactttg gttcaaacaa gatgataagt tttttttgcc gaaggcttgt
     2641 ctcaactttg aatttttcag cccatttgct tatgtggacc ccttgcactg taacatggcc
     2701 tatttgtacc ttgagctcct caaagactca ctcaacgagt atgcatatgc agcagagcta
     2761 gcaggcttga gctatgatct ccaaaatacc atctatggga tgtatctttc agtgaaaggt
     2821 tacaatgaca agcagccaat tttactaaag aagattattg agaaaatggc tacctttgag
     2881 attgatgaaa aaagatttga aattatcaaa gaagcatata tgcgatctct taacaatttc
     2941 cgggctgaac agcctcacca gcatgccatg tactacctcc gcttgctgat gactgaagtg
     3001 gcctggacta aagatgagtt aaaagaagct ctggatgatg taacccttcc tcgccttaag
     3061 gccttcatac ctcagctcct gtcacggctg cacattgaag cccttctcca tggaaacata
     3121 acaaagcagg ctgcattagg aattatgcag atggttgaag acaccctcat tgaacatgct
     3181 cataccaaac ctctccttcc aagtcagctg gttcggtata gagaagttca gctccctgac
     3241 agaggatggt ttgtttatca gcagagaaat gaagttcaca ataactgtgg catcgagata
     3301 tactaccaaa cagacatgca aagcacctca gagaatatgt ttctggagct cttctgtcag
     3361 attatctcgg aaccttgctt caacaccctg cgcaccaagg agcagttggg ctatatcgtc
     3421 ttcagcgggc cacgtcgagc taatggcata cagggcttga gattcatcat ccagtcagaa
     3481 aagccacctc actacctaga aagcagagtg gaagctttct taattaccat ggaaaagtcc
     3541 atagaggaca tgacagaaga ggccttccaa aaacacattc aggcattagc aattcgtcga
     3601 ctagacaaac caaagaagct atctgctgag tgtgctaaat actggggaga aatcatctcc
     3661 cagcaatata attttgacag agataacact gaggttgcat atttaaagac acttaccaag
     3721 gaagatatca tcaaattcta caaggaaatg ttggcagtag atgctccaag gagacataag
     3781 gtatccgtcc atgttcttgc cagggaaatg gattcttgtc ctgttgttgg agagttccca
     3841 tgtcaaaatg acataaattt gtcacaagca ccagccttgc cacaacctga agtgattcag
     3901 aacatgaccg aattcaagcg tggtctgcca ctgtttcccc ttgtgaaacc acatattaac
     3961 ttcatggctg caaaactcga attcgccgcc accatggcgg ccgccagctg gagccaccct
     4021 cagttcgaga agggaggagg aagcggcgga ggcagcggag gaggaagctg gagccacccg
     4081 cagttcgaga aatgactcga gcatgcatct agaggctgga gccacccgca gttcgagaaa
     4141 tgactcgagc atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     4201 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4261 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4321 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4381 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4441 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4501 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4561 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4621 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4681 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4741 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4801 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4861 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4921 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4981 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5041 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5101 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5161 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5221 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5281 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5341 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5401 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5461 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5521 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5581 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5641 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5701 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5761 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5821 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5881 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5941 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6001 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6061 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6121 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6181 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6241 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     6301 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6361 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6421 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6481 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6541 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6661 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7021 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7261 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8041 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8281 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8461 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8521 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8581 ttccgcgcac atttccccga aaagtgcca
//
