LOCUS       40924_42709        6888 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTc6HN, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6888)
  AUTHORS   Batista M, Marchini FK, Celedon PA, Fragoso SP, Probst CM, Preti H, 
            Ozaki LS, Buck GA, Goldenberg S, Krieger MA.
  TITLE     A high-throughput cloning system for reverse genetics in Trypanosoma
            cruzi
  JOURNAL   BMC Microbiol. 10 (1), 259 (2010)
  PUBMED    20942965
REFERENCE   2  (bases 1 to 6888)
  AUTHORS   Batista M, Marchini FK, Celedon PAF., Fragoso SP, Probst CM, Preti 
            H, Ozaki LS, Buck GA, Goldenberg S, Krieger MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-APR-2010) Instituto Carlos Chagas, Professor Algacyr 
            Munhoz Mader, 3775, Curitiba, Parana 81350010, Brasil
REFERENCE   3  (bases 1 to 6888)
  AUTHORS   Batista M, Marchini FK, Celedon PAF., Fragoso SP, Probst CM, Preti 
            H, Ozaki LS, Buck GA, Goldenberg S, Krieger MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-NOV-2010) Instituto Carlos Chagas, Professor Algacyr 
            Munhoz Mader, 3775, Curitiba, Parana 81350010, Brasil
REFERENCE   4  (bases 1 to 6888)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6888)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Microbiol."; date: "2010"; volume: "10"; issue: "1"; pages: "259"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-APR-2010) Instituto Carlos Chagas, Professor Algacyr Munhoz 
            Mader, 3775, Curitiba, Parana 81350010, Brasil"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (18-NOV-2010) Instituto Carlos Chagas, Professor Algacyr Munhoz 
            Mader, 3775, Curitiba, Parana 81350010, Brasil"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     On Nov 18, 2010 this sequence version replaced HM162846.1.
FEATURES             Location/Qualifiers
     source          1..6888
                     /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..1720
                     /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             3600..4388
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     promoter        complement(4700..4718)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4739..4755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4763..4779)
                     /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(4787..4817)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4832..4853)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5141..5729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5903..6760)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6761..6865)
                     /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 agtcactatg 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 tctaccatag
     3181 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
     3241 taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt
     3301 gatactagtt agtctgcgat gctgtggacc gacgcattga aatacacacc gtctttggcg
     3361 ttcctttttt tttttttgat ttgttttttt tattgagatg ttttgttttg tttttttcag
     3421 tttttattat atcagcagtt tgtccgcctg cattcatgca gtgtttggta attctttcta
     3481 ttttttggaa ttatggcgat aatattcttg tctttaaact tcttataacc aattgtgctt
     3541 tagagtttcc tgcttagttg ctattaacac actgttagga acgtgaaacc ctcgagatga
     3601 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3661 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3721 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3781 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3841 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3901 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3961 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4021 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4081 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgaatg cccgacggcg
     4141 atgatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4201 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4261 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4321 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4381 agttcttctg actcgagtct gcgatgctgt ggaccgacgc attgaaatac acaccgtctt
     4441 tggcgttcct tttttttttt ttgatttgtt ttttttattg agatgttttg ttttgttttt
     4501 ttcagttttt attatatcag cagtttgtcc gcctgcattc atgcagtgtt tggtaattct
     4561 ttctattttt tggaattatg gcgataatat tcttgtcttt aaacttctta taaccaattg
     4621 tgctttagag tttcctgctt agttgctatt aacacactgt taggaacgtg aaaccgggcc
     4681 cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     4741 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4801 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4861 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4921 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4981 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5041 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5101 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5161 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5221 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5281 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5341 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5401 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5461 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5521 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5581 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5641 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5701 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5761 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5821 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5881 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5941 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6001 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6061 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6121 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6181 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6241 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6301 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6361 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6421 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6481 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6541 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6601 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6661 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6721 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6781 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6841 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
