LOCUS       40924_42704        6477 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Protein fusion vector pTc6HisP-NH 1.1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6477)
  AUTHORS   Kugeratski FG, Batista M, Inoue AH, Ramos BD, Krieger MA, Marchini/ 
            FK.
  TITLE     pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma 
            cruzi gene characterisation
  JOURNAL   Mem. Inst. Oswaldo Cruz (2015) In press
  PUBMED    26200713
REFERENCE   2  (bases 1 to 6477)
  AUTHORS   Kugeratski FG, Batista M, Inoue AH, Ramos BD, Krieger MA, Marchini 
            FK.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2015) Mass Spectrometry Platform, Instituto Carlos
            Chagas, Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 
            81350010, Brasil
REFERENCE   3  (bases 1 to 6477)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6477)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mem. Inst. 
            Oswaldo Cruz (2015) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-APR-2015) Mass Spectrometry Platform, Instituto Carlos Chagas, 
            Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 81350010, 
            Brasil"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6477
                     /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      659..1275
                     /note="derived from Trypanosoma cruzi 18S ribosomal RNA
                     gene promoter region"
                     /regulatory_class="promoter"
     CDS             1569..1586
                     /label=6xHis
                     /note="6xHis affinity tag"
     protein_bind    1596..1715
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1745..1775
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1829..2485
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2830..3132
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3176..3300)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3609..3980
                     /label=BleoR
                     /note="antibiotic-binding protein"
     promoter        complement(4289..4307)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4328..4344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4352..4368)
                     /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(4376..4406)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4421..4442)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4730..5318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5492..6349)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6350..6454)
                     /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 tggagctcca
      661 ccagtttcct tgcggggctt atttggtgtg ttggattttt aattttttga tgattcggag
      721 gaattttttg caatttttgg gtacccctaa aaaaattttt ggagtttccg gggttcgatc
      781 caacttgtgc ttttttaacc gtccatttgt gtctagtaca tcatggagcg gtattctctc
      841 cggtggccca ggtggcgtcg gaaggaattg gtgtccctca ggatgtttgg tttttcgttt
      901 ttttttttgc ggtcttttgt ttcgcatcgc cgtatcgtgt gcggcgcgac cttctttttg
      961 gctcattggt cagtgaggtt tataataaag tggatcattt gggtttttgg ccctttgcgg
     1021 cgcacccgcg gaatggggga gggattttca tataatgtag tgtacattgt gtgtgggtcg
     1081 caaaatgtca tgccgtgtgc aatgtttatt tgcgtgctgt gcgtgtcctt ttgtgattcc
     1141 accacttgtg tgtattttac cttatcgtct ctccctgccg ttggctgtgc gtgaattgtg
     1201 cttcgcattc gtgggaaatc aaaaggggaa tcaacgtgtt cttttttttc agaaggggag
     1261 tcaaagcgcc gtgtggatat ctctgcgatg ctgtggaccg acgcattgaa atacacaccg
     1321 tctttggcgt tccttttttt ttttttgatt tgtttttttt attgagatgt tttgttttgt
     1381 ttttttcagt ttttattata tcagcagttt gtccgcctgc attcatgcag tgtttggtaa
     1441 ttctttctat tttttggaat tatggcgata atattcttgt ctttaaactt cttataacca
     1501 attgtgcttt agagtttcct gcttagttgc tattaacaca ctgttaggaa cgtgaaacca
     1561 tggcatgcca tcaccatcac catcactcta gaatcacaag tttgtacaaa aaagctgaac
     1621 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga
     1681 ctacataata ctgtaaaaca caacatatcc agtcatattg gcggccgcat taggcacccc
     1741 aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc
     1801 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     1861 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     1921 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     1981 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     2041 ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     2101 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     2161 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     2221 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     2281 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     2341 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     2401 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     2461 ctgcgatgag tggcagggcg gggcgtaaac gcgtggatcc ggcttactaa aagccagata
     2521 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     2581 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     2641 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     2701 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     2761 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     2821 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     2881 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     2941 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     3001 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     3061 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     3121 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag
     3181 tgactggata tgttgtgttt tacagcatta tgtagtctgt tttttatgca aaatctaatt
     3241 taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt
     3301 gatactagtc cggcgcgcca ttgatctgcg atgctgtgga ccgacgcatt gaaatacaca
     3361 ccgtctttgg cgttcctttt tttttttttg atttgttttt tttattgaga tgttttgttt
     3421 tgtttttttc agtttttatt atatcagcag tttgtccgcc tgcattcatg cagtgtttgg
     3481 taattctttc tattttttgg aattatggcg ataatattct tgtctttaaa cttcttataa
     3541 ccaattgtgc tttagagttt cctgcttagt tgctattaac acactgttag gaacgtgaaa
     3601 ccaccggtat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     3661 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     3721 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     3781 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     3841 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     3901 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     3961 tcgtggccga ggagcaggac ctcgagtctg cgatgctgtg gaccgacgca ttgaaataca
     4021 caccgtcttt ggcgttcctt tttttttttt tgatttgttt tttttattga gatgttttgt
     4081 tttgtttttt tcagttttta ttatatcagc agtttgtccg cctgcattca tgcagtgttt
     4141 ggtaattctt tctatttttt ggaattatgg cgataatatt cttgtcttta aacttcttat
     4201 aaccaattgt gctttagagt ttcctgctta gttgctatta acacactgtt aggaacgtga
     4261 aaccgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     4321 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4381 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4441 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4501 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4561 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4621 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4681 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4741 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4801 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5101 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5161 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5221 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5281 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5341 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5401 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5461 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5521 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5581 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5641 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5701 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5761 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5821 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5881 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5941 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6001 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6061 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6121 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6181 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6241 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6301 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6361 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6421 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
