LOCUS       62056_21020        7428 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcFlagN-NH 2.0, complete sequence.
ACCESSION   MT180301
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7428)
  AUTHORS   Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
  TITLE     Characterising ISWI chromatin remodeler in Trypanosoma cruzi
  JOURNAL   Mem. Inst. Oswaldo Cruz 115, e190457 (2020)
  PUBMED    32428081
REFERENCE   2  (bases 1 to 7428)
  AUTHORS   Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2020) LACTAS, Fiocruz, Professor Algacyr Munhoz 
            Mader, Curitiba 81350010, Brasil
REFERENCE   3  (bases 1 to 7428)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mem. Inst. 
            Oswaldo Cruz 115, e190457 (2020)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-MAR-2020) LACTAS, Fiocruz, Professor Algacyr Munhoz Mader, 
            Curitiba 81350010, Brasil"
FEATURES             Location/Qualifiers
     source          1..7428
                     /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"
     regulatory      706..1322
                     /note="Trypanosoma cruzi 18S ribosomal RNA promoter region"
                     /regulatory_class="promoter"
     misc_feature    1335..1839
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511469.50 CDS"
     CDS             1849..2643
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    2653..2970
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511145.40 CDS"
     CDS             2986..3051
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     protein_bind    3061..3185
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3210..3240
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3294..3950
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4295..4597
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4641..4765)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4789..5209
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     promoter        complement(5240..5258)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5279..5295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5303..5319)
                     /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(5327..5357)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5372..5393)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5681..6269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6443..7300)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7301..7405)
                     /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 tggagctcgc
      661 ggccgcccac ttgtatcgtg ggcgccactt gaatcgtgga tcgatcacca gtttccttgc
      721 ggggcttatt tggtgtgttg gatttttaat tttttgatga ttcggaggaa ttttttgcaa
      781 tttttgggta cccctaaaaa aatttttgga gtttccgggg ttcgatccaa cttgtgcttt
      841 tttaaccgtc catttgtgtc tagtacatca tggagcggta ttctctccgg tggcccaggt
      901 ggcgtcggaa ggaattggtg tccctcagga tgtttggttt ttcgtttttt tttttgcggt
      961 cttttgtttc gcatcgccgt atcgtgtgcg gcgcgacctt ctttttggct cattggtcag
     1021 tgaggtttat aataaagtgg atcatttggg tttttggccc tttgcggcgc acccgcggaa
     1081 tgggggaggg attttcatat aatgtagtgt acattgtgtg tgggtcgcaa aatgtcatgc
     1141 cgtgtgcaat gtttatttgc gtgctgtgcg tgtccttttg tgattccacc acttgtgtgt
     1201 attttacctt atcgtctctc cctgccgttg gctgtgcgtg aattgtgctt cgcattcgtg
     1261 ggaaatcaaa aggggaatca acgtgttctt ttttttcaga aggggagtca aagcgccgtg
     1321 tgatcgatca tatggatttg ggtaggatga gaagggagtt gctgaggaaa aggggaggga
     1381 atcattaata attagaagag ttttttcttc tctctcattt cctgtatcta tatatgtatt
     1441 aggagctgta ttggcaactt ggtggagcat cggcggggtg tttgcggcgc ggacctctgc
     1501 aaacttccaa aggtggtaat gggaatttga aataacttgc caatggacat gactcaacag
     1561 acgaaagcgg tagcaacgat actatgtaat agctgttgtt gttgttattt ttatgtttat
     1621 ttttattatt gatacttcgc ttcggtactc tctctgtttt tttttttctc tcactcattc
     1681 cctctttata tatatatata tatatatata tatatatata tatcgtacgt gtctttttgt
     1741 tgttgttctc aacttcacgt tggctttttt tttttttttt ttgacttgga ttgtatcgga
     1801 cggcccaata acaataataa ttaaaataaa agtggggaaa agcttactag tatgattgaa
     1861 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     1921 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     1981 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     2041 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     2101 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     2161 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     2221 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     2281 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     2341 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gaatgcccga cggcgatgat
     2401 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     2461 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     2521 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     2581 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     2641 ttctgactcg agcggcatcg gatgcgtttt atggttctgt cttttttttc ccctcctttt
     2701 tttttttctt ttaatttatt ttactttact gcacggtacc ttgtaggaaa tgagatgtgt
     2761 tttttttttt gttttttttg ttgttgttgt tgttgttgag attttttttt tttttttttt
     2821 ttttgatctt acgtgtgtcc tgtgttactg agaatattta tgctgcgcat ttggagatat
     2881 tcatagttgt ttaaatggac tttttctgct gctcaaactt ttgttttctg cgtaattaac
     2941 agggaaggaa actttctaag tcgttcatca gatatctcta gaatggacta caaggatcac
     3001 gacggcgatt acaaggacca cgacattgat tacaaggacg atgacgataa ggcatgcatc
     3061 acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta
     3121 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
     3181 ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
     3241 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
     3301 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     3361 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     3421 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     3481 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     3541 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     3601 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     3661 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     3721 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     3781 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     3841 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     3901 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaacgcgtg
     3961 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     4021 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     4081 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     4141 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     4201 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     4261 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     4321 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     4381 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     4441 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     4501 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     4561 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     4621 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     4681 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     4741 gttcagcttt cttgtacaaa gtggtgatat aagcggccgc ggcgcgccta tggtagtgaa
     4801 tggatatata tatatatata tatatatata tataaggtgt aaatgaaatt agtggaagag
     4861 ggaattagat gggtagcgcg ttgcgtggcg ctggggagtg tctcgtttta tgacttttgt
     4921 atctctctgt ttgagtgtgt gtgtgtgtgt gtgtgtgttt tattcttttt ttgttgttgt
     4981 tgttcttgtt aaatatttat taatgatttg cttttccttc ttgtgtggct tgaaattatt
     5041 tcctctactg tttgtctcca cgtcaatttt gcccataact catcttgact ttcaatcctt
     5101 agcgagtgtt ttgttgtttt tgggatagtc gttttttgtt tgtgtgtgtg tgtgtgtgtg
     5161 tgtgtatttt tttctttttt cttcttcttg gaggggctcc tacacgttta ccggtgggcc
     5221 cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     5281 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5341 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5401 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6121 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
