LOCUS       sequence-171480-        4039 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  P. falciparum plasmid for expression of both diCre fragments from 
            single casette using skip peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4039)
  AUTHORS   Birnbaum J, Flemming S, Reichard N, Soares AB, Mesen-Ramirez P, 
            Jonscher E, Bergmann B, Spielmann T
  TITLE     A genetic system to study Plasmodium falciparum protein function.
  JOURNAL   Nat Methods. 2017 Mar 13. doi: 10.1038/nmeth.4223.
  PUBMED    28288121
REFERENCE   2  (bases 1 to 4039)
  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: "Nat 
            Methods. 2017 Mar 13. doi: 10.1038/nmeth.4223."
COMMENT     Sequence Label: pSkipFlox
FEATURES             Location/Qualifiers
     source          1..4039
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             152..550
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(1130..1149)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1131..1149)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1156..1173)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1156..1172)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1165..1187)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1381..1400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1500..1522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1560..1578)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1646..1750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1751..1819,1820..2611)
                     /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     complement(1969..1988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2782..3370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3271..3290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3524..3541
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3658..3679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3694..3699,3700..3717,3718..3724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3732..3748
                     /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)."
     primer_bind     3737..3759
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3756..3772
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3756..3772
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3790..3808
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3790..3807
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 attcttgtca caataaaaat atatatatat atatatatat ttataatatg tatattttaa
       61 actagaaaag gaataactaa tattttattt attatcattc aagatttata ttttataata
      121 ataaatacct aatagaaata tatcaggatc catggccaag cctttgtctc aagaagaatc
      181 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
      241 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
      301 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
      361 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
      421 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
      481 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
      541 ggagggctaa aagcttattt aataatagat taaaaatatt ataaaaataa aaacataaac
      601 acagaaatta caaaaaaaat acatatgaat ttttttttgt aatcttcctt ataaatatag
      661 aataatgaat catataaaac atatcattat tcatttattt acatttaaaa ttattgtttc
      721 agtatcttta atttattatg tatatataaa aataacttac aattttatta ataagcaata
      781 tatgtttatt aattcatgtt ttgtaattta tgggatagcg atttttttta ctgtctgtat
      841 ttttcttttt taattatgtt ttaattgtat tttattttta ttattgttct ttttatagta
      901 ttattttaaa acaaaatgta ttttctaaga acttataata ataataaata taaattttaa
      961 taaaaattat atttatcttt tacaatatga acataaagta caacattaat atatagcttt
     1021 taatattttt attcctaatc atgtaaatct taaatttttc tttttaaaca tatgttaaat
     1081 atttatttct cattatatat aagaacatat ttattaaatc tagaattcgc cctatagtga
     1141 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     1201 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     1261 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     1321 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     1381 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     1441 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     1501 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     1561 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1621 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1741 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1801 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1861 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1921 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1981 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2041 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2101 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2161 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2221 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2281 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2341 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2401 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2461 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2521 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2581 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2641 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2701 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3361 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3421 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3481 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3541 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3601 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3661 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     3721 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     3781 cgccaagcta tttaggtgac actatagaat actcaagctg cggccgctaa cgtaacagac
     3841 ttaggaggag atcttagtag ttgagtgatt ctatatacat atacataaat aaattacata
     3901 atattataaa ttttttttat attatattag aatgtttact atataaaaat aaatattttc
     3961 ttatatattt ttttattttt ccatatgaaa aaagaaattt tttatatatt tttttattat
     4021 attttataaa taaaaaaga
//
