LOCUS       62056_2970        5505 bp DNA     circular SYN 15-JUL-2020
DEFINITION  Cloning vector pALH015, complete sequence.
ACCESSION   MT345687
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5505)
  AUTHORS   Herneisen AL, Sidik SM, Markus BM, Drewry DH, Zuercher WJ, Lourido 
            S.
  TITLE     Identifying the Target of an Antiparasitic Compound in Toxoplasma 
            Using Thermal Proteome Profiling
  JOURNAL   ACS Chem. Biol. (2020) In press
  PUBMED    32597628
REFERENCE   2  (bases 1 to 5505)
  AUTHORS   Herneisen AL, Sidik SM, Markus BM, Zuercher WJ, Lourido S.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-APR-2020) Whitehead Institute, Massachusetts Institute
            of Technology, 455 Main St, Cambridge, MA 02142, USA
REFERENCE   3  (bases 1 to 5505)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Chem. 
            Biol. (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-APR-2020) Whitehead Institute, Massachusetts Institute of 
            Technology, 455 Main St, Cambridge, MA 02142, USA"
FEATURES             Location/Qualifiers
     source          1..5505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     5'UTR           698..1195
                     /label=TgTUB1 5'UTR
                     /note="TgTUB1 5'UTR"
     CDS             1196..2545
                     /label=GCaMP6f
                     /note="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
     3'UTR           2549..3291
                     /label=3'UTR of DHFR
                     /note="3'UTR of DHFR"
     promoter        complement(3317..3335)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3356..3372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3380..3396)
                     /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)."
     promoter        complement(3404..3434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3449..3470)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3758..4346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4520..5377)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5378..5482)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgatat taattaacat gcatgtcccg cgttcgtgaa
      721 attctctgca tcagcggagt gatcaggaat catcgtctca gcgggatgac gttgcggagc
      781 aggccggctc gcggtgggca gtcagatgcc gaaggcgtaa ctcaggacgg cttgcgctca
      841 tcgcagaaca ggggtggtgc ctgcattggg tgcggttggt gatcctggtt ggaccggtgg
      901 agatgcgcgc gcacgaaggg gatgtgtcag aaacattttg tttgttctct gtgaactttt
      961 agatgtgtta aaggcggcga atattagcag agagtcctcc ttgttggatt ctctcttgaa
     1021 tttcgccctt tccttctctt tgcgagtctc gtagagaaca agcactcgtt cgccgtccct
     1081 gacgacgcaa cccgcgcaga agacatccac caaacggtgt tacacaatca ccttgtgtga
     1141 agttcttgcg gaaaactact cgttggcatt ttttcttgaa ttcccttttt cgacaatggg
     1201 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
     1261 ggatctgtac gacgatgacg ataaggatct cgccaccatg gtcgactcat cacgtcgtaa
     1321 gtggaataag acaggtcacg cagtcagagc tataggtcgg ctgagctcac tcgagaacgt
     1381 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     1441 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1501 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     1561 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1621 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga
     1681 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1741 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa
     1801 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1861 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1921 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     1981 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2041 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacct
     2101 gccggaccaa ctgactgaag agcagatcgc agaatttaaa gaggaattct ccctatttga
     2161 caaggacggg gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtccctggg
     2221 gcagaacccc acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga
     2281 cggcacaatc gacttccctg agttcctgac aatgatggca agaaaaatga aatacaggga
     2341 cacggaagaa gaaattagag aagcgttcgg tgtgtttgat aaggatggca atggctacat
     2401 cagtgcagca gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga
     2461 ggttgatgaa atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga
     2521 gtttgtacaa atgatgacag cgaagtaaac agaagctgcc cgtctctcgt tttcctctct
     2581 tttcggaggg atcagggaga gtgcctcggg tcggagagag ctgacgaggg ggtgccagag
     2641 acccctgtgt cctttatcga agaaaaggga tgactcttca tgtggcattt cacacagtct
     2701 cacctcgcct tgttttcttt ttgtcaatca gaacgaaagc gagttgcggg tgacgcagat
     2761 gtgcgtgtat ccactcgnga atgcgttatc gttctgtatg ccgctagagt gctggactgt
     2821 tgctgtctgc ccacgacagc agacaacttt ccttctatgc acttgcagga tggtgcagcg
     2881 caaacgacgg agagaaagga gcaccctctc agtttcccta cgatgtgctg tcagtttcga
     2941 ctcttcaccg cgaacgattg gcgatacgtc tctgttgact tgttaggctc cgaccacgaa
     3001 gctcccttaa ctagataagc cgcgacacct aagtgtacac catttgcaga tcgataatct
     3061 gcgaccgctg aatccgtcca gatcagtaaa accgcaccac ctaagtgtaa accttgttta
     3121 ggtcgataaa atgctaccaa cccccaccca caatcgagcc ttgagcgttt ctgcgcacgc
     3181 gttggcctac gtgacttgct gatgcctgcc tctggccatt catgccagtc agtgcgcata
     3241 aaaatgtgga cacagtcggt tgacaagtgt tctggcaggc tacagtgaca cgcggccgca
     3301 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     3361 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3421 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3481 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3541 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3601 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3661 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3721 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3781 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3841 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3901 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3961 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4021 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4081 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4141 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4201 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4261 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4321 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4381 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4441 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4561 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4621 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4681 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4741 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4801 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4861 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4921 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4981 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5041 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5101 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5161 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5221 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5281 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5341 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5401 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5461 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
