LOCUS       V016107                 6526 bp    DNA     circular SYN 11-FEB-2020
DEFINITION  Exported.
ACCESSION   V016107
VERSION     V016107
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6526)
  AUTHORS   Swift RP, Rajaram K, Liu HB, Dziedzic A, Jedlicka AE, Roberts AD,
            Matthews KA, Jhun H, Bumpus NN, Tewari SG, Wallqvist A, Prigge ST.
  TITLE     A mevalonate bypass system facilitates elucidation of plastid
            biology in malaria parasites
  JOURNAL   PLoS Pathog. (2020) In press
REFERENCE   2  (bases 1 to 6526)
  AUTHORS   Prigge ST.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-DEC-2019) Molecular Microbiology and Immunology, Johns
            Hopkins SPH, 615 N. Wolfe, rm E5132, Johns Hopkins SPH, Baltimore,
            MD 21205, USA
REFERENCE   3  (bases 1 to 6526)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS
            Pathog. (2020) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (11-DEC-2019) Molecular Microbiology and Immunology, Johns Hopkins
            SPH, 615 N. Wolfe, rm E5132, Johns Hopkins SPH, Baltimore, MD 21205,
            USA"
FEATURES             Location/Qualifiers
     source          1..6526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_structure  9..30
                     /note="NotI LIC site; ligation-independent cloning"
     5'UTR           37..1018
                     /label="CAM promoter"
                     /note="CAM promoter"
     CDS             1025..1585
                     /gene="DHFR"
                     /label="Dihydrofolate reductase"
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     3'UTR           1591..2155
                     /label="HRPII 3'"
                     /note="HRPII 3'"
     misc_structure  2165..2186
                     /note="NgoMIV LIC site; ligation-independent cloning"
     5'UTR           2191..3340
                     /label="U6 snRNA 5'"
                     /note="U6 snRNA 5'"
     misc_feature    3342..3347
                     /label="BsaI endonuclease site"
                     /note="BsaI endonuclease site"
     protein_bind    3354..3375
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3390..3420
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3428..3444
                     /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)."
     CDS             3464..3637
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     terminator      complement(3709..3752)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    3754..3759
                     /label="BsaI endonuclease site"
                     /note="BsaI endonuclease site"
     misc_RNA        3761..3836
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     3'UTR           3837..4343
                     /label="U6 snRNA 3'"
                     /note="U6 snRNA 3'"
     promoter        4628..4732
                     /label="AmpR promoter"
     CDS             4733..5590
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5764..6352
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 gcggccgtgc cacgagcggc cgctgcgctt aagcattttg taaaaaaaat taaaatatat
       61 ttatataata ttattttatt ttattatata ttatattatt tttattttta tttttatttt
      121 tttttctcta caaattttat ctattggttt attataaaaa tatctatttc taataataaa
      181 taattaagat atcaatttat agaaacaaaa tatatacttg tataatttta tttttttata
      241 taaatcatta catatataat tatacaatat tttttctaag agataattat atattaatat
      301 atataaaaaa aggtgttttt tttttttttt tttattttta tttttatttt atggtaatat
      361 tttattttcc ttattttata aattatatta gtttatatgt gattaatttt atatattatc
      421 aatttatata tttttaaatg cttacttaat tatctttttt tttttttttt tttttttttc
      481 ccctcttttt atattaattt atttttgaaa aaattgatat atatatatat atataatata
      541 tatatataca tgtagtagta ttaaacaatg tataatatat ataaataata tatttatata
      601 tttcatttca attttaattt tttttggttt tttttttttt tctttttgtc atatttaaaa
      661 aaaattatat tcatataagt tatgcatttt ttataaacat tattcaatat atgtataata
      721 taatatatat atatatatta atgtattatt ccaatgtgca tgataaaaga aaaaaataat
      781 atttataaaa aaaaagaaaa ataaaacaaa aaaagaaaaa aaaaaaaaaa aaaaaaaaaa
      841 tacaaaaata aataatataa tttataatta tatattcttg tcacaataaa aatatatata
      901 tatatatata tatttataat atgtatattt taaactagaa aaggaataac taatatttta
      961 tttattatca ttcaagattt atattttata ataataaata cctaatagaa atatatcagg
     1021 atccatgcat ggttcgctaa actgcatcgt cgctgtgtcc cagaacatgg gcatcggcaa
     1081 gaacggggac tacccctggc caccgctcag gaacgaattt agatatttcc agagaatgac
     1141 cacaacctct tcagtagaag gtaaacagaa tctggtgatt atgggtaaga agacctggtt
     1201 ctccattcct gagaagaatc gacctttaaa gggtagaatt aatttagttc tcagcagaga
     1261 actcaaggaa cctccacaag gagctcattt tctttccaga agtctagatg atgccttaaa
     1321 acttactgaa caaccagaat tagcaaataa agtagacatg gtctggatag ttggtggcag
     1381 ttctgtttat aaggaagcca tgaatcaccc aggccatctt aaactatttg tgacaaggat
     1441 catgcaagac tttgaaagtg acacgttttt tccagaaatt gatttggaga aatataaact
     1501 tctgccagaa tacccaggtg ttctctctga tgtccaggag gagaaaggca ttaagtacaa
     1561 atttgaagta tatgagaaga atgattaagc ttatttaata atagattaaa aatattataa
     1621 aaataaaaac ataaacacag aaattacaaa aaaaatacat atgaattttt tttttgtaat
     1681 cttccttata aatatagaat aatgaatcat ataaaacata tcattattca tttatttaca
     1741 tttaaaatta ttgtttcagt atctttaatt tattatgtat atataaaaat aacttacaat
     1801 tttattaata aacaatatat gtttattaat tcatgttttg taatttatgg gatagcgatt
     1861 ttttttactg tctgtatttt tcttttttaa ttatgtttta attgtatttt atttttatta
     1921 ttgttctttt tatagtatta ttttaaaaca aaatgtattt tctaagaact tataataata
     1981 ataaatataa attttaataa aaattatatt tatcttttac aatatgaaca taaagtacaa
     2041 cattaatata tagcttttaa tatttttatt cctaatcatg taaatcttaa atttttcttt
     2101 ttaaacatat gttaaatatt tatttctcat tatatataag aacatattta ttaaatctag
     2161 aattcgacag acgccggctg gtggccatgg aacggtaaaa ataataacac gaattatcac
     2221 aataattttt cacatatgta tatatatata tatattatca aagggacaac cgtttcaaga
     2281 aagcaaatgt tattattttg tttttattct aatgtatgta agccaaaaac caataaaaaa
     2341 gtaataatta aaaaggaaat aaaataaata tacatatgta attatactta tataataagt
     2401 atgtatattt tttattattt ttttttttta atgctattta tttattattg catcttttta
     2461 tttaattttt gtattttttt atataattcc tttatttctt tatttctttc tttatttctt
     2521 tttttttttt tttttttttt tttttttcct ttaattaaac tttaaaagac gttataaatt
     2581 ttaaacatat tttataaata aaaaaaaaaa aaaaaaagac gaaataataa aataaacgaa
     2641 gataaagtaa tattttgtta ttatgtcaca tagtatgtgt actacaatta tttatggatg
     2701 caaattatta aattggtagt tattatatac atacatgttt ttttttttgt gtgtattaaa
     2761 aggaactatt atataaatag aacaatttca tgacatttgg atttctacac atcttgaggt
     2821 tttacaataa tatcttcttt tttttttttt ctcttctata tatgaaatat attgtaaaaa
     2881 ggtacaatat atataatatt tatatactat gaaaaaaaaa aaatatatac ataaactaat
     2941 aataatttat tctaatttaa tataaaaaaa aattcttgct tgttcagata aataaaagaa
     3001 atttatataa atagctatat tttttttttt tttttttttt tttttttcta attataaata
     3061 taattattca tttaaaaaat tttaaatatt taaatatttc ataaattaaa aaaaaaaaaa
     3121 aaaaaactat taatataata aacttttatt tttactgtaa tataattttt tataatgtaa
     3181 aaataaaggg taaattatta ttaaaaaatg tatatgttat gtatatataa cataatatat
     3241 tataatatat atatatatat atatatatat ataatattag agtaaccaaa atgcataatt
     3301 tttcctatat gcacatattt catattaagt atataatatt tgagacccgc aattaatgtg
     3361 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     3421 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga
     3481 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3541 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3601 tcgcccttcc caacagttgc gcagtctgaa tggcgaatgg cagcttggct gttttggcgg
     3661 atgagataag attttcagcc tgatacagat taaatcagaa cgcagaagat aaaacgaaag
     3721 gcccagtcga aagactgggc ctttcgtttt atcggtctca gttttagagc tagaaatagc
     3781 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     3841 ttattatttc ctataaaata atatatatat atatatatat atatatatat atatatttta
     3901 aatataatga agtttttaag gctaccataa aaaaaaaaat acatatatat ataaataaat
     3961 atattatatt atataatata tatattaatg tttttgtttt ttaatatttt tttttttttt
     4021 tgtaaaatat attatttttg gacttataaa taattgtgta catacatata ttattttatt
     4081 ataataaata taataatact ataataatct ttatgcaata tttgtaattc atatataata
     4141 atttttaaaa ttatctatct tttgaaaaaa aaaaaaaaat aaaaataata aaaattataa
     4201 tgttattaaa tactataatt attatgatta atataatata tatatatata tatatatata
     4261 tatatatata tatatatttt atatcattta atatattatg tgtgatataa taataaatcc
     4321 attgtccata aatagatgaa atagcatatg gtgcactctc agtacaatct gctctgatgc
     4381 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4441 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4501 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4561 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5281 aatgccaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggtt ctcgcggtat
     5461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5881 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatcga aaaacgccag
     6361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6421 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6481 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaa
//
