LOCUS       sequence-382978-        3815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector that expresses the EDHFR protein with 
            FKBP12 fused to it.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3815)
  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 3815)
  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-EDHFR-FKBP12
FEATURES             Location/Qualifiers
     source          1..3815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             28..501
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             532..855
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    924..1058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(961..980)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1015..1034
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1097..1114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1268..1856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1348..1367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2027..2818,2819..2887))
                     /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     2650..2669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2888..2992)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3032..3052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(3037..3366,3367..3381,3382..3650)
                     /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      3217..3352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             3382..3650
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          3680..3776
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 aattccacca tgggcgggtc gagcggaatc agtctgattg cggcgttagc ggtagatcgc
       61 gttatcggca tggaaaacgc catgccgtgg aacctgcctg ccgatctcgc ctggtttaaa
      121 cgcaacacct taaataaacc cgtgattatg ggccgccata cctgggaatc aatcggtcgt
      181 ccgttgccag gacgcaaaaa tattatcctc agcagtcaac cgggtacgga cgatcgcgta
      241 acgtgggtga agtcggtgga tgaagccatc gcggcgtgtg gtgacgtacc agaaatcatg
      301 gtgattggcg gcggtcgcgt ttatgaacag ttcttgccaa aagcgcaaaa actgtatctg
      361 acgcatatcg acgcagaagt ggaaggcgac acccatttcc cggattacga gccggatgac
      421 tgggaatcgg tattcagcga attccacgat gctgatgcgc agaactctca cagctattgc
      481 tttgagattc tggagcggcg gggcagcggc catatgggcg ggtcgctgca gggagtgcag
      541 gtggaaacca tctccccggg agacgggcgc accttcccca agcgcggcca gacctgcgtg
      601 gtgcactaca ccgggatgct tgaagatgga aagaaatttg attcctcccg ggacagaaac
      661 aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt
      721 gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt
      781 gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt
      841 ctaaaactgg aataatctag agaaaaaacc tcccacacct ccccctgaac ctgaaacata
      901 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
      961 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1021 tgtccaaact catcaatgta tcttatcatg tctggatccc cgggtaccga gctcgaatta
     1081 attcctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     1141 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1201 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1261 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1321 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1381 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1441 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1501 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1561 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1621 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1681 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1741 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1801 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2161 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     2221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2341 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2641 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2881 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3001 cccgaaaagt gccacctgca agcttggctg tggaatgtgt gtcagttagg gtgtggaaag
     3061 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3121 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3181 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3241 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3301 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3361 tgcaaaaagc tcctcgatcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc
     3421 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc
     3481 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg
     3541 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt
     3601 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctctga
     3661 aaaaccagaa agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc
     3721 cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct
     3781 gtacggaagt gttacttctg ctctaaaagc tgcgg
//
