LOCUS       sequence-382976-        4025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector that expresses the EGFP protein with 
            FKBP12 fused to it.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4025)
  AUTHORS   Bogaert A, Van de Steene T, Vuylsteke M, Eyckerman S, Gevaert K
  TITLE     A decoupled Virotrap approach to study the interactomes of 
            N-terminal proteoforms.
  JOURNAL   Methods Enzymol. 2023;684:253-287. doi: 10.1016/bs.mie.2023.02.003. 
            Epub 2023 Mar 10.
  PUBMED    37230591
REFERENCE   2  (bases 1 to 4025)
  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: "Methods 
            Enzymol."; date: "2023"; volume: "684:253-287. doi"; pages: " 
            10.1016/bs.mie.2023.02.003. Epub 2023 Mar 10"
COMMENT     Sequence Label: pMET7-EGFP-FKBP12
FEATURES             Location/Qualifiers
     source          1..4025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..33)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(18..347,348..362,363..631)
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early 
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     rep_origin      198..333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             363..631
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          661..757
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(806..808,809..811,812..1522)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(806..808,809..811,812..1522)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(851..872)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1112..1131)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1459..1480
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1538..1861
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    1930..2064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1967..1986)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2021..2040
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2103..2120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2274..2862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2354..2373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3033..3824,3825..3893))
                     /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     3656..3675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3894..3998)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
       61 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
      181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
      241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
      301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgatc
      361 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc
      421 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct
      481 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac
      541 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg
      601 tttcgttttc tgttctgcgc cgttacagat ccaagctctg aaaaaccaga aagttaactg
      661 gtaagtttag tctttttgtc ttttatttca ggtcccggat ccggtggtgg tgcaaatcaa
      721 agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggaag tgttacttct
      781 gctctaaaag ctgcggaatt ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      841 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      901 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      961 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1021 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1081 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1141 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1201 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1261 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     1321 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1381 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     1441 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1501 cggcatggac gagctgtaca agggcgggtc gctgcaggga gtgcaggtgg aaaccatctc
     1561 cccgggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg
     1621 gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc cctttaagtt
     1681 tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt
     1741 gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc
     1801 aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa aactggaata
     1861 atctagagaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     1921 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1981 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2041 aatgtatctt atcatgtctg gatccccggg taccgagctc gaattaattc ctcttccgct
     2101 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2161 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2221 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2281 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2341 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2401 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2461 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2521 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2581 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2641 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2701 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2761 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2821 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2881 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2941 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3001 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3061 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3121 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3181 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3241 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3301 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3361 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3421 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3481 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3541 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3601 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3661 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3721 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3781 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3841 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3901 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3961 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4021 cctgc
//
