LOCUS       40924_2333        8685 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p-REX2mDHFR-int, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8685)
  AUTHORS   Mesen-Ramirez P, Reinsch F, Blancke Soares A, Bergmann B, Ullrich 
            AK, Tenzer S, Spielmann T.
  TITLE     Stable Translocation Intermediates Jam Global Protein Export in 
            Plasmodium falciparum Parasites and Link the PTEX Component EXP2 
            with Translocation Activity
  JOURNAL   PLoS Pathog. 12 (5), E1005618 (2016)
  PUBMED    27168322
REFERENCE   2  (bases 1 to 8685)
  AUTHORS   Blancke Soares A, Spielmann T.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2016) Parasitology Section, Bernhard Nocht 
            Institute for Tropical Medicine, Bernhard-Nocht-Str. 74, Hamburg 
            20359, Germany
REFERENCE   3  (bases 1 to 8685)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8685)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2016"; volume: "12"; issue: "5"; pages: "E1005618"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2016) Parasitology Section, Bernhard Nocht Institute for 
            Tropical Medicine, Bernhard-Nocht-Str. 74, Hamburg 20359, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      36..1499
                     /label=Pfcrt 5' region
                     /note="Pfcrt 5' region"
                     /regulatory_class="promoter"
     misc_feature    1506..1787
                     /note="similar to REX2; P. falciparum ring exported protein
                     2 (PF3D7_0936000)"
     misc_feature    1794..2354
                     /note="similar to mDHFR; mouse dihydrofolate reductase"
     CDS             1794..2354
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     CDS             2361..3074
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      3089..3541
                     /label=P. berghei DT 3' region
                     /note="P. berghei DT 3' region"
                     /regulatory_class="terminator"
     regulatory      complement(3553..3898)
                     /label=PfHOP 5' region
                     /note="PfHOP 5' region"
                     /regulatory_class="promoter"
     regulatory      3899..4389
                     /label=PfCAM 5' region
                     /note="PfCAM 5' region"
                     /regulatory_class="promoter"
     CDS             4396..4956
                     /gene="DHFR"
                     /label=DHFR
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     CDS             4966..5019
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5026..5421
                     /label=BSD
                     /note="blasticidin S deaminase"
     regulatory      5429..5998
                     /label=PfhrpII 3' region
                     /note="PfhrpII 3' region"
                     /regulatory_class="terminator"
     promoter        complement(6003..6020)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6027..6043)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6517..6621
                     /label=AmpR promoter
     CDS             6622..7479
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7653..8241
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8529..8550
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8565..8595
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8603..8619
                     /label=lac operator
                     /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)."
     primer_bind     8627..8643
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8661..8679
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gcggccgcta acgtaacaga cttaggagga gatcttagta gttgagtgat tctatataca
       61 tatacataaa taaattacat aatattataa atttttttta tattatatta gaatgtttac
      121 tatataaaaa taaatatttt cttatatatt tttttatttt ttcatatgaa aaaagaaatt
      181 ttttatatat ttttttatta tattttataa ataaaaaaga acttatacct attattatta
      241 ttatatataa aaaatatata tttttttata aattcctatt tttccgatta tttatttatt
      301 tttttttttt attgtttaaa aatatataaa aaaaattctt atatatttaa tattataagt
      361 ccataaatat atataattta tataatttat atatttccat acttttttta ttatatatca
      421 tgagataaaa taaatttcat atagaaatcg tatttattta ttaatttgat atatattaat
      481 aaataaatat taataataat atgttatata tatatatatt tatttaatta ttatatagga
      541 aactatatat atatatatat attatatttt tttttttgaa atataataat atataatatt
      601 attcctaaaa atatctatat tcttatacct gaaccctttt tttttttttt tttttttttt
      661 gactttcgat tgttcattct gtttattgta taaatatata aatatatata tattgtatat
      721 tttattacat atattttttt tttttgaaag ttaagaattt taacttaata aaaaaagtac
      781 atatattttt aataaatgtc ctccattata taaattgtta tataaaagat tttatataat
      841 ttaaatagaa ttcatttata aacaaatttg tttataaaaa tatatttatg tatatataat
      901 ataaatatat atatttatat atatatatat atatatatta ctatatatat tttttttttt
      961 ttttcctttt ttttactttc ccaagttgta ctgcttctaa gcttttttaa taaacatata
     1021 taatttgtac aaatatttta gattatatac atgatgtata tttgaatata ttttctatat
     1081 atttgtggtt ccatttttgt atattatata taatatattt atatatatat tgatatgtca
     1141 atatttgtat aacacatgaa gtttttgttt tttttttttt ttttttttaa tggagaatat
     1201 ttaataatat atgaaaaaaa ttttatataa atatatatat atatatatat atatatatat
     1261 atatatatat atatatgtat atatatatat ttatatatac atgtatgttt tttaaaaagt
     1321 taaataattc tatagattat tttcattgtc ttcacatata tgacataaat attttaaaat
     1381 cgacattccg atatattata tttttagact ataatatccg ttaataataa atacacgcag
     1441 tcatattatt tattatacat tcatttatta ttttgttttt tttaatttct tacatataac
     1501 tcgagatgaa aatgtattta gctgaaattt ttagttctgg taaagagtct ttgttatctt
     1561 taaaggatac tttaggatct agtaattttt ctccactgaa gccatgtgaa ggccttgagt
     1621 gtttaccaca agtattattt ttgtacgtta tatttctttt attatgtaca ggaattttta
     1681 tgcataataa gaataagctt aaaaaacaac aaaggttaac aaatatgaat gattcatatg
     1741 atgcaaaaaa ttcaggattc gcagaacagt atgatcaaca acaacatcct aggatggttc
     1801 gaccattgaa ctgcatcgtc gccgtgtccc aaaatatggg gattggcaag aacggagacc
     1861 taccctggcc tccgctcagg aacgagttca agtacttcca aagaatgacc acaacctctt
     1921 cagtggaagg taaacagaat ctggtgatta tgggtaggaa aacctggttc tccattcctg
     1981 agaagaatcg acctttaaag gacagaatta atatagttct cagtagagaa ctcaaagaac
     2041 caccacgagg agctcatttt cttgccaaaa gtttggatga tgccttaaga cttattgaac
     2101 aaccggaatt ggcaagtaaa gtagacatgg tttggatagt cggaggcagt tctgtttacc
     2161 aggaagccat gaatcaacca ggccacctca gactctttgt gacaaggatc atgcaggaat
     2221 ttgaaagtga cacgtttttc ccagaaattg atttggggaa atataaactt ctcccagaat
     2281 acccaggcgt cctctctgag gtccaggagg aaaaaggcat caagtataag tttgaagtct
     2341 acgagaagaa agacggtacc atgagtaaag gagaagaact tttcactgga gttgtcccaa
     2401 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg
     2461 aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac
     2521 ctgttccatg gccaacactt gtcactactt tcgcgtatgg tcttcaatgc tttgcgagat
     2581 acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac
     2641 aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt
     2701 ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg
     2761 gaaacattct tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg
     2821 cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg
     2881 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     2941 ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa
     3001 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
     3061 atgaactata caaataaccc gggtcgaggg atatggcagc ttaatgttcg tttttcttat
     3121 ttatatattt ataccaattg attgtattta taactgtaaa aatgtgtatg ttgtgtgcat
     3181 attttttttt gtgcatgcac atgcatgtaa atagctaaaa ttatgaacat tttatttttt
     3241 gttcagaaaa aaaaaacttt acacacataa aatggctagt atgaatagcc atattttata
     3301 taaattaaat cctatgaatt tatgaccata ttaaaaattt agatatttat ggaacataat
     3361 atgtttgaaa caataagaca aaattattat tattattatt atttttactg ttataattat
     3421 gtgtctcctt caatgattca taaatagttg gacttgattt ttaaaatgtt tataatatga
     3481 ttagcatagt taaataaaaa aagttgaaaa attaaaaaaa aacatataaa cacaaatgat
     3541 ggtttttcct tcaatttcga tatcaattta tagaaacaaa atatatactt gtataatttt
     3601 atttttttat ataaatcatt acatatataa ttatacaata ttttttctaa gagataatta
     3661 tatattaata tatataaaaa aaggtgtttt tttttttttt ttttattttt atttttattt
     3721 tatggtaata ttttattttc cttattttat aaattatatt agtttatatg tgattaattt
     3781 tatatattat caatttatat atttttaaat gcttacttaa ttatcttttt tttttttttt
     3841 tttttttttt cccctctttt tatattaatt tatttttgaa aaaattgata tatatatata
     3901 tatataatat atatatatac atgtagtagt attaaacaat gtataatata tataaataat
     3961 atatttatat atttcatttc aattttaatt ttttttggtt tttttttttt ttctttttgt
     4021 catatttaaa aaaaattata ttcatataag ttatgcattt tttataaaca ttattcaata
     4081 tatgtataat ataatatata tatatatatt aatgtattat tccaatgtgc atgataaaag
     4141 aaaaaaataa tatttataaa aaaaaagaaa aataaaacaa aaaaagaaaa aaaaaaaaaa
     4201 aaaaaaaaaa atacaaaaat aaataatata atttataatt atatattctt gtcacaataa
     4261 aaatatatat atatatatat atatttataa tatgtatatt ttaaactaga aaaggaataa
     4321 ctaatatttt atttattatc attcaagatt tatattttat aataataaat acctaataga
     4381 aatatatcac tgcagatgca tggttcgcta aactgcatcg tcgctgtgtc ccagaacatg
     4441 ggcatcggca agaacgggga ctacccctgg ccaccgctca ggaacgaatt tagatatttc
     4501 cagagaatga ccacaacctc ttcagtagaa ggtaaacaga atctggtgat tatgggtaag
     4561 aagacctggt tctccattcc tgagaagaat cgacctttaa agggtagaat taatttagtt
     4621 ctcagcagag aactcaagga acctccacaa ggagctcatt ttctttccag aagtctagat
     4681 gatgccttaa aacttactga acaaccagaa ttagcaaata aagtagacat ggtctggata
     4741 gttggtggca gttctgttta taaggaagcc atgaatcacc caggccatct taaactattt
     4801 gtgacaagga tcatgcaaga ctttgaaagt gacacgtttt ttccagaaat tgatttggag
     4861 aaatataaac ttctgccaga atacccaggt gttctctctg atgtccagga ggagaaaggc
     4921 attaagtaca aatttgaagt atatgagaag aatgatgcta gcggagaagg aagaggaagt
     4981 ttattaacat gtggagatgt agaagaaaat ccaggaccag gatccatggc caagcctttg
     5041 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     5101 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     5161 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     5221 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     5281 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     5341 gcgatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     5401 tctggttatg tgtgggaggg ctaaccgctt atttaataat agattaaaaa tattataaaa
     5461 ataaaaacat aaacacagaa attacaaaaa aaatacatat gaattttttt tttgtaatct
     5521 tccttataaa tatagaataa tgaatcatat aaaacatatc attattcatt tatttacatt
     5581 taaaattatt gtttcagtat ctttaattta ttatgtatat ataaaaataa cttacaattt
     5641 tattaataaa caatatatgt ttattaattc atgttttgta atttatggga tagcgatttt
     5701 ttttactgtc tgtatttttc ttttttaatt atgttttaat tgtattttat ttttattatt
     5761 gttcttttta tagtattatt ttaaaacaaa atgtattttc taagaactta taataataat
     5821 aaatataaat tttaataaaa attatattta tcttttacaa tatgaacata aagtacaaca
     5881 ttaatatata gcttttaata tttttattcc taatcatgta aatcttaaat ttttcttttt
     5941 aaacatatgt taaatattta tttctcatta tatataagaa catatttatt aaatctagaa
     6001 ttctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     6061 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6121 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6181 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     6241 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     6301 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     6361 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     6421 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6481 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6541 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6601 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6661 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6721 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6781 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6841 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6901 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6961 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7021 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7081 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7141 gacgagcgtg acaccacgat gcctgtagca atgccaacaa cgttgcgcaa actattaact
     7201 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7261 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7321 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7381 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7441 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7501 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7561 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7621 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7681 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7741 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     7801 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7861 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7921 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7981 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     8041 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8101 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8161 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8221 ggggggcgga gcctatcgaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8281 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8341 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8401 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8461 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8521 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8581 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8641 gaccatgatt acgccaagct atttaggtga cactatagaa tactc
//
