LOCUS       62056_21040        7461 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcHAN-NH 2.0, complete sequence.
ACCESSION   MT180299
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7461)
  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 7461)
  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 7461)
  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..7461
                     /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..3075
                     /label=3xHA
                     /note="three tandem HA epitope tags"
     protein_bind    3094..3218
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3243..3273
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3327..3983
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4328..4630
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4674..4798)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4822..5242
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     promoter        complement(5273..5291)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5312..5328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5336..5352)
                     /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(5360..5390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5405..5426)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5714..6302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6476..7333)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7334..7438)
                     /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 gaatgtaccc gtacgacgtg
     3001 cccgactatg cgggctatcc gtatgacgtc ccggactatg caggttccta tccgtatgac
     3061 gttccagatt acgctccggc cgccgcatgc atcacaagtt tgtacaaaaa agctgaacga
     3121 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     3181 acataatact gtaaaacaca acatatccag tcactatggc ggccgcatta ggcaccccag
     3241 gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag gatccgtcga
     3301 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     3361 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     3421 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     3481 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     3541 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     3601 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     3661 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     3721 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     3781 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     3841 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     3901 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     3961 gcgatgagtg gcagggcggg gcgtaaacgc gtggatccgg cttactaaaa gccagataac
     4021 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     4081 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
     4141 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     4201 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     4261 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
     4321 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     4381 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
     4441 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     4501 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     4561 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     4621 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc gaccatagtg
     4681 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     4741 atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac aaagtggtga
     4801 tataagcggc cgcggcgcgc ctatggtagt gaatggatat atatatatat atatatatat
     4861 atatataagg tgtaaatgaa attagtggaa gagggaatta gatgggtagc gcgttgcgtg
     4921 gcgctgggga gtgtctcgtt ttatgacttt tgtatctctc tgtttgagtg tgtgtgtgtg
     4981 tgtgtgtgtg ttttattctt tttttgttgt tgttgttctt gttaaatatt tattaatgat
     5041 ttgcttttcc ttcttgtgtg gcttgaaatt atttcctcta ctgtttgtct ccacgtcaat
     5101 tttgcccata actcatcttg actttcaatc cttagcgagt gttttgttgt ttttgggata
     5161 gtcgtttttt gtttgtgtgt gtgtgtgtgt gtgtgtgtat ttttttcttt tttcttcttc
     5221 ttggaggggc tcctacacgt ttaccggtgg gcccggtacc cagcttttgt tccctttagt
     5281 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5341 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5401 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5461 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5521 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5581 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5641 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5701 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5761 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5821 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5881 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5941 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6001 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6061 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6121 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6181 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6241 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6301 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6361 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6421 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6481 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6541 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6601 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6661 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6721 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6781 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6841 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6901 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6961 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7021 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7081 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7141 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7201 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7261 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7321 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7381 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7441 catttccccg aaaagtgcca c
//
