LOCUS       62056_21065        7386 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcStrepN-CO 2.0, complete sequence.
ACCESSION   MT180302
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      665..1281
                     /note="Trypanosoma cruzi 18S ribosomal RNA promoter region"
                     /regulatory_class="promoter"
     misc_feature    1294..1798
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511469.50 CDS"
     protein_bind    1816..1940
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1965..1995
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2049..2705
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             3050..3352
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3396..3520)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3534..3557
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     misc_feature    3575..3995
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     CDS             4005..4799
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    4809..5126
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511145.40 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 tggagctcat
      661 cgatcaccag tttccttgcg gggcttattt ggtgtgttgg atttttaatt ttttgatgat
      721 tcggaggaat tttttgcaat ttttgggtac ccctaaaaaa atttttggag tttccggggt
      781 tcgatccaac ttgtgctttt ttaaccgtcc atttgtgtct agtacatcat ggagcggtat
      841 tctctccggt ggcccaggtg gcgtcggaag gaattggtgt ccctcaggat gtttggtttt
      901 tcgttttttt ttttgcggtc ttttgtttcg catcgccgta tcgtgtgcgg cgcgaccttc
      961 tttttggctc attggtcagt gaggtttata ataaagtgga tcatttgggt ttttggccct
     1021 ttgcggcgca cccgcggaat gggggaggga ttttcatata atgtagtgta cattgtgtgt
     1081 gggtcgcaaa atgtcatgcc gtgtgcaatg tttatttgcg tgctgtgcgt gtccttttgt
     1141 gattccacca cttgtgtgta ttttacctta tcgtctctcc ctgccgttgg ctgtgcgtga
     1201 attgtgcttc gcattcgtgg gaaatcaaaa ggggaatcaa cgtgttcttt tttttcagaa
     1261 ggggagtcaa agcgccgtgt gatcgatcat atggatttgg gtaggatgag aagggagttg
     1321 ctgaggaaaa ggggagggaa tcattaataa ttagaagagt tttttcttct ctctcatttc
     1381 ctgtatctat atatgtatta ggagctgtat tggcaacttg gtggagcatc ggcggggtgt
     1441 ttgcggcgcg gacctctgca aacttccaaa ggtggtaatg ggaatttgaa ataacttgcc
     1501 aatggacatg actcaacaga cgaaagcggt agcaacgata ctatgtaata gctgttgttg
     1561 ttgttatttt tatgtttatt tttattattg atacttcgct tcggtactct ctctgttttt
     1621 ttttttctct cactcattcc ctctttatat atatatatat atatatatat atatatatat
     1681 atcgtacgtg tctttttgtt gttgttctca acttcacgtt ggcttttttt tttttttttt
     1741 tgacttggat tgtatcggac ggcccaataa caataataat taaaataaaa gtggggaaaa
     1801 gcttgcggcc gcatcacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata
     1861 aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca
     1921 caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt
     1981 ccggctcgta taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg
     2041 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     2101 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     2161 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     2221 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
     2281 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     2341 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     2401 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     2461 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     2521 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     2581 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     2641 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     2701 gggcgtaaac gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg
     2761 ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag
     2821 gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa
     2881 ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt
     2941 cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg
     3001 tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa
     3061 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     3121 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     3181 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     3241 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     3301 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     3361 aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt
     3421 tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat
     3481 cattttacgt ttctcgttca gctttcttgt acaaagtggt gatgtctaga atgtggagcc
     3541 acccgcagtt cgaaaaggca tgctaaggcg cgcctatggt agtgaatgga tatatatata
     3601 tatatatata tatatatata aggtgtaaat gaaattagtg gaagagggaa ttagatgggt
     3661 agcgcgttgc gtggcgctgg ggagtgtctc gttttatgac ttttgtatct ctctgtttga
     3721 gtgtgtgtgt gtgtgtgtgt gtgttttatt ctttttttgt tgttgttgtt cttgttaaat
     3781 atttattaat gatttgcttt tccttcttgt gtggcttgaa attatttcct ctactgtttg
     3841 tctccacgtc aattttgccc ataactcatc ttgactttca atccttagcg agtgttttgt
     3901 tgtttttggg atagtcgttt tttgtttgtg tgtgtgtgtg tgtgtgtgtg tatttttttc
     3961 ttttttcttc ttcttggagg ggctcctaca cgtttaccgg tactagtatg attgaacaag
     4021 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4081 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4141 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4201 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4261 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4321 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4381 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4441 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4501 tcgcgccagc cgaactgttc gccaggctca aggcgcgaat gcccgacggc gatgatctcg
     4561 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4621 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4681 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4741 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4801 gactcgagcg gcatcggatg cgttttatgg ttctgtcttt tttttcccct cctttttttt
     4861 tttcttttaa tttattttac tttactgcac ggtaccttgt aggaaatgag atgtgttttt
     4921 ttttttgttt tttttgttgt tgttgttgtt gttgagattt tttttttttt tttttttttt
     4981 gatcttacgt gtgtcctgtg ttactgagaa tatttatgct gcgcatttgg agatattcat
     5041 agttgtttaa atggactttt tctgctgctc aaacttttgt tttctgcgta attaacaggg
     5101 aaggaaactt tctaagtcgt tcatcagata tcccacttgt atcgtgggcg ccacttgaat
     5161 cgtgggcggc cgcgggcccg 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
//
