LOCUS       62056_21035        7461 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcHAN-CO 2.0, complete sequence.
ACCESSION   MT180298
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      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..3623
                     /label=3xHA
                     /note="three tandem HA epitope tags"
     misc_feature    3650..4070
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     CDS             4080..4874
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    4884..5201
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511145.40 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 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 atgtacccgt
     3541 acgacgtgcc cgactatgcg ggctatccgt atgacgtccc ggactatgca ggttcctatc
     3601 cgtatgacgt tccagattac gctccggccg ccgcatgcta aggcgcgcct atggtagtga
     3661 atggatatat atatatatat atatatatat atataaggtg taaatgaaat tagtggaaga
     3721 gggaattaga tgggtagcgc gttgcgtggc gctggggagt gtctcgtttt atgacttttg
     3781 tatctctctg tttgagtgtg tgtgtgtgtg tgtgtgtgtt ttattctttt tttgttgttg
     3841 ttgttcttgt taaatattta ttaatgattt gcttttcctt cttgtgtggc ttgaaattat
     3901 ttcctctact gtttgtctcc acgtcaattt tgcccataac tcatcttgac tttcaatcct
     3961 tagcgagtgt tttgttgttt ttgggatagt cgttttttgt ttgtgtgtgt gtgtgtgtgt
     4021 gtgtgtattt ttttcttttt tcttcttctt ggaggggctc ctacacgttt accggtacta
     4081 gtatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4141 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4201 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4261 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4321 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4381 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4441 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4501 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4561 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgaatgcccg
     4621 acggcgatga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4681 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4741 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4801 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4861 ttgacgagtt cttctgactc gagcggcatc ggatgcgttt tatggttctg tctttttttt
     4921 cccctccttt ttttttttct tttaatttat tttactttac tgcacggtac cttgtaggaa
     4981 atgagatgtg tttttttttt tgtttttttt gttgttgttg ttgttgttga gatttttttt
     5041 tttttttttt tttttgatct tacgtgtgtc ctgtgttact gagaatattt atgctgcgca
     5101 tttggagata ttcatagttg tttaaatgga ctttttctgc tgctcaaact tttgttttct
     5161 gcgtaattaa cagggaagga aactttctaa gtcgttcatc agatatccca cttgtatcgt
     5221 gggcgccact tgaatcgtgg gcggccgcgg 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
//
