LOCUS       62056_21070        7386 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcStrepN-NH 2.0, complete sequence.
ACCESSION   MT180303
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7386)
  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 7386)
  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 7386)
  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..7386
                     /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..3009
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     protein_bind    3019..3143
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3168..3198
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3252..3908
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4253..4555
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4599..4723)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4747..5167
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     promoter        complement(5198..5216)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5237..5253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5261..5277)
                     /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(5285..5315)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5330..5351)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5639..6227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6401..7258)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7259..7363)
                     /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 gaatgtggag ccacccgcag
     3001 ttcgaaaagg catgcatcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat
     3061 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     3121 acacaacata tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg
     3181 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
     3241 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     3301 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     3361 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     3421 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
     3481 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     3541 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     3601 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     3661 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     3721 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     3781 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     3841 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     3901 gcggggcgta aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc
     3961 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     4021 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     4081 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     4141 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     4201 cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt
     4261 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     4321 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     4381 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     4441 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     4501 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     4561 gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg
     4621 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     4681 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggtgatataa gcggccgcgg
     4741 cgcgcctatg gtagtgaatg gatatatata tatatatata tatatatata taaggtgtaa
     4801 atgaaattag tggaagaggg aattagatgg gtagcgcgtt gcgtggcgct ggggagtgtc
     4861 tcgttttatg acttttgtat ctctctgttt gagtgtgtgt gtgtgtgtgt gtgtgtttta
     4921 ttcttttttt gttgttgttg ttcttgttaa atatttatta atgatttgct tttccttctt
     4981 gtgtggcttg aaattatttc ctctactgtt tgtctccacg tcaattttgc ccataactca
     5041 tcttgacttt caatccttag cgagtgtttt gttgtttttg ggatagtcgt tttttgtttg
     5101 tgtgtgtgtg tgtgtgtgtg tgtatttttt tcttttttct tcttcttgga ggggctccta
     5161 cacgtttacc ggtgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
     5221 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5281 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5341 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5401 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5461 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5521 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5581 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5641 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5701 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5761 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5821 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5881 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5941 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6001 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6061 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6121 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6181 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6241 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6301 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6361 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6421 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6481 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6541 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6601 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6661 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6721 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6781 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6841 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6901 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6961 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7021 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7081 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7141 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7201 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7261 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7321 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7381 tgccac
//
