LOCUS       62056_21075        7392 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcTy1N-CO 2.0, complete sequence.
ACCESSION   MT180308
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      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"
     misc_feature    3531..3572
                     /label=Ty1 epitope tag
                     /note="Ty1 epitope tag"
     misc_feature    3581..4001
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     CDS             4011..4805
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    4815..5132
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511145.40 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 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 atggaggtgc
     3541 acacgaacca ggacccgctg gacgcatgct aaggcgcgcc tatggtagtg aatggatata
     3601 tatatatata tatatatata tatataaggt gtaaatgaaa ttagtggaag agggaattag
     3661 atgggtagcg cgttgcgtgg cgctggggag tgtctcgttt tatgactttt gtatctctct
     3721 gtttgagtgt gtgtgtgtgt gtgtgtgtgt tttattcttt ttttgttgtt gttgttcttg
     3781 ttaaatattt attaatgatt tgcttttcct tcttgtgtgg cttgaaatta tttcctctac
     3841 tgtttgtctc cacgtcaatt ttgcccataa ctcatcttga ctttcaatcc ttagcgagtg
     3901 ttttgttgtt tttgggatag tcgttttttg tttgtgtgtg tgtgtgtgtg tgtgtgtatt
     3961 tttttctttt ttcttcttct tggaggggct cctacacgtt taccggtact agtatgattg
     4021 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4081 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4141 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4201 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4261 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4321 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4381 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4441 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4501 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgaatgccc gacggcgatg
     4561 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4621 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4681 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4741 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4801 tcttctgact cgagcggcat cggatgcgtt ttatggttct gtcttttttt tcccctcctt
     4861 tttttttttc ttttaattta ttttacttta ctgcacggta ccttgtagga aatgagatgt
     4921 gttttttttt ttgttttttt tgttgttgtt gttgttgttg agattttttt tttttttttt
     4981 ttttttgatc ttacgtgtgt cctgtgttac tgagaatatt tatgctgcgc atttggagat
     5041 attcatagtt gtttaaatgg actttttctg ctgctcaaac ttttgttttc tgcgtaatta
     5101 acagggaagg aaactttcta agtcgttcat cagatatccc acttgtatcg tgggcgccac
     5161 ttgaatcgtg ggcggccgcg 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
//
