LOCUS       62056_21015        7428 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcFlagN-CO 2.0, complete sequence.
ACCESSION   MT180300
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7428)
  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 7428)
  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 7428)
  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..7428
                     /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..3599
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     misc_feature    3617..4037
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     CDS             4047..4841
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    4851..5168
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511145.40 CDS"
     promoter        complement(5240..5258)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5279..5295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5303..5319)
                     /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(5327..5357)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5372..5393)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5681..6269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6443..7300)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7301..7405)
                     /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 atggactaca
     3541 aggatcacga cggcgattac aaggaccacg acattgatta caaggacgat gacgataagg
     3601 catgctaagg cgcgcctatg gtagtgaatg gatatatata tatatatata tatatatata
     3661 taaggtgtaa atgaaattag tggaagaggg aattagatgg gtagcgcgtt gcgtggcgct
     3721 ggggagtgtc tcgttttatg acttttgtat ctctctgttt gagtgtgtgt gtgtgtgtgt
     3781 gtgtgtttta ttcttttttt gttgttgttg ttcttgttaa atatttatta atgatttgct
     3841 tttccttctt gtgtggcttg aaattatttc ctctactgtt tgtctccacg tcaattttgc
     3901 ccataactca tcttgacttt caatccttag cgagtgtttt gttgtttttg ggatagtcgt
     3961 tttttgtttg tgtgtgtgtg tgtgtgtgtg tgtatttttt tcttttttct tcttcttgga
     4021 ggggctccta cacgtttacc ggtactagta tgattgaaca agatggattg cacgcaggtt
     4081 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4141 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4201 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4261 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4321 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4381 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4441 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4501 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4561 tcgccaggct caaggcgcga atgcccgacg gcgatgatct cgtcgtgacc catggcgatg
     4621 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4681 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4741 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4801 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgactcgag cggcatcgga
     4861 tgcgttttat ggttctgtct tttttttccc ctcctttttt tttttctttt aatttatttt
     4921 actttactgc acggtacctt gtaggaaatg agatgtgttt ttttttttgt tttttttgtt
     4981 gttgttgttg ttgttgagat tttttttttt tttttttttt ttgatcttac gtgtgtcctg
     5041 tgttactgag aatatttatg ctgcgcattt ggagatattc atagttgttt aaatggactt
     5101 tttctgctgc tcaaactttt gttttctgcg taattaacag ggaaggaaac tttctaagtc
     5161 gttcatcaga tatcccactt gtatcgtggg cgccacttga atcgtgggcg gccgcgggcc
     5221 cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     5281 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5341 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5401 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6121 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
