LOCUS       62056_21080        7392 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcTy1N-NH 2.0, complete sequence.
ACCESSION   MT180309
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7392)
  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 7392)
  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 7392)
  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..7392
                     /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"
     misc_feature    2983..3015
                     /label=Ty1 epitope tag
                     /note="Ty1 epitope tag"
     protein_bind    3025..3149
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3174..3204
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3258..3914
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4259..4561
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4605..4729)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4753..5173
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     promoter        complement(5204..5222)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5243..5259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5267..5283)
                     /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(5291..5321)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5336..5357)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5645..6233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6407..7264)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7265..7369)
                     /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 gaatggaggt gcacacgaac
     3001 caggacccgc tggacgcatg catcacaagt ttgtacaaaa aagctgaacg agaaacgtaa
     3061 aatgatataa atatcaatat attaaattag attttgcata aaaaacagac tacataatac
     3121 tgtaaaacac aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac
     3181 tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag
     3241 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     3301 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     3361 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     3421 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta
     3481 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     3541 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     3601 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     3661 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     3721 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     3781 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     3841 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     3901 ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt
     3961 atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt
     4021 caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca
     4081 gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat
     4141 gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag
     4201 gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat
     4261 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     4321 gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct
     4381 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     4441 gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc
     4501 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     4561 ataaatgtca ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat
     4621 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
     4681 tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtg atataagcgg
     4741 ccgcggcgcg cctatggtag tgaatggata tatatatata tatatatata tatatataag
     4801 gtgtaaatga aattagtgga agagggaatt agatgggtag cgcgttgcgt ggcgctgggg
     4861 agtgtctcgt tttatgactt ttgtatctct ctgtttgagt gtgtgtgtgt gtgtgtgtgt
     4921 gttttattct ttttttgttg ttgttgttct tgttaaatat ttattaatga tttgcttttc
     4981 cttcttgtgt ggcttgaaat tatttcctct actgtttgtc tccacgtcaa ttttgcccat
     5041 aactcatctt gactttcaat ccttagcgag tgttttgttg tttttgggat agtcgttttt
     5101 tgtttgtgtg tgtgtgtgtg tgtgtgtgta tttttttctt ttttcttctt cttggagggg
     5161 ctcctacacg tttaccggtg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     5221 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5281 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5341 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5401 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5461 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5521 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5821 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6061 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7381 gaaaagtgcc ac
//
