LOCUS       sequence-417473-        6440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FKBP10 fused with EGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6440)
  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 6440)
  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: FKBP10-EGFP
FEATURES             Location/Qualifiers
     source          1..6440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2375..2384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2381..2383,2384..2386,2387..3100)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2426..2447)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2687..2706)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3034..3055
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3223..3344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3260..3279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3314..3333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3351..3806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3488..3509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3700..3719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3833..3937
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3939..4296
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3939..4130
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3962..3982)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4147..4282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4331..5125
                     /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             4331..5125
                     /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(4385..4404)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4995..5014
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5313..5332)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5357..5404
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5733..6321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6222..6241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga
      601 ctcagatctc gagatgttcc ccgcgggccc ccccagccac agcctcctcc ggctccccct
      661 gctgcagttg ctgctactgg tggtgcaggc cgtggggagg gggctgggcc gcgccagccc
      721 ggccgggggc cccctggaag atgtggtcat cgagaggtac cacatcccca gggcctgtcc
      781 ccgggaagtg cagatggggg attttgtgcg ctaccactac aacggcactt ttgaagatgg
      841 caagaagttt gattcaagct atgatcgcaa caccttggtg gccatcgtgg tgggtgtggg
      901 gcgcctcatc actggcatgg accgaggcct catgggcatg tgtgtcaacg agcggcgacg
      961 cctcattgtg cctccccacc tgggctatgg gagcatcggc ctggcggggc tcattccacc
     1021 ggatgccacc ctctacttcg atgtggttct gctggatgtg tggaacaagg aagacaccgt
     1081 gcaggtgagc acattgctgc gcccgcccca ctgcccccgc atggtccagg acggcgactt
     1141 tgtccgctac cactacaatg gcaccctgct ggacggcacc tccttcgaca ccagctacag
     1201 taagggcggc acttatgaca cctacgtcgg ctctggttgg ctgatcaagg gcatggacca
     1261 ggggctgctg ggcatgtgtc ctggagagag aaggaagatt atcatccctc cattcctggc
     1321 ctatggcgag aaaggctatg ggacagtgat ccccccacag gcctcgctgg tctttcacgt
     1381 cctcctgatt gacgtgcaca acccgaagga cgctgtccag ctagagacgc tggagctccc
     1441 ccccggctgt gtccgcagag ccggggccgg ggacttcatg cgctaccact acaatggctc
     1501 cttgatggac ggcaccctct tcgattccag ctactcccgc aaccacacct acaataccta
     1561 tatcgggcag ggttacatca tccccgggat ggaccagggg ctgcagggtg cctgcatggg
     1621 ggaacgccgg agaattacca tccccccgca cctcgcctat ggggagaatg gaactggaga
     1681 caagatccct ggctctgccg tgctaatctt caacgtccat gtcattgact tccacaaccc
     1741 tgcggatgtg gtggaaatca ggacactgtc ccggccatct gagacctgca atgagaccac
     1801 caagcttggg gactttgttc gataccatta caactgttct ttgctggacg gcacccagct
     1861 gttcacctcg catgactacg gggcccccca ggaggcgact ctcggggcca acaaggtgat
     1921 cgaaggcctg gacacgggcc tgcagggcat gtgtgtggga gagaggcggc agctcatcgt
     1981 gcccccgcac ctggcccacg gggagagtgg agcccgggga gtcccaggca gtgctgtgct
     2041 gctgtttgag gtggagctgg tgtcccggga ggatgggctg cccacaggct acctgtttgt
     2101 gtggcacaag gaccctcctg ccaacctgtt tgaagacatg gacctcaaca aggatggcga
     2161 ggtccctccg gaggagttct ccaccttcat caaggctcaa gtgagtgagg gcaaaggacg
     2221 cctcatgcct gggcaggacc ctgagaaaac cataggagac atgttccaga accaggaccg
     2281 caaccaggac ggcaagatca cagtcgacga gctcaagctg aagtcagatg aggacgagga
     2341 gcgggtccac gaggagctcg cggatccacc ggtcgccacc atggtgagca agggcgagga
     2401 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2461 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2521 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2581 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2641 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2701 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2761 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2821 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2881 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2941 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     3001 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3061 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc
     3121 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3181 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3241 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3301 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3361 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3421 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3481 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3541 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3601 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3661 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3721 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3781 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3841 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3901 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3961 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4021 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4141 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4201 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4261 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4321 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4381 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4441 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4501 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4561 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4621 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4681 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4741 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4801 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4861 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4921 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4981 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5041 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5101 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5161 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5221 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5281 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5341 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5401 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5461 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5521 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     5581 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5641 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5701 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5761 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5821 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6241 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6301 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6361 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6421 attaccgcca tgcattagtt
//
