LOCUS       40924_42784        6906 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTcMYCN, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  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 6906)
  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 6906)
  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 6906)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6906)
  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 HM162841.1.
FEATURES             Location/Qualifiers
     source          1..6906
                     /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"
     misc_feature    1569..1604
                     /label=c-myc epitope tag
                     /note="c-myc epitope tag"
     protein_bind    1614..1733
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1763..1793
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1847..2503
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2848..3150
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3194..3318)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3618..4406
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     promoter        complement(4718..4736)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4757..4773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4781..4797)
                     /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(4805..4835)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4850..4871)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5159..5747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5921..6778)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6779..6883)
                     /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 tggcatgcat ggaacaaaag ttgatttctg aatttgaatt gaattctaga atcacaagtt
     1621 tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
     1681 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcatattggc
     1741 ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggat
     1801 tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     1861 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     1921 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     1981 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     2041 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
     2101 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     2161 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     2221 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     2281 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     2341 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     2401 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     2461 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaaacgc gtggatccgg
     2521 cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat
     2581 atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt
     2641 acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct
     2701 ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga
     2761 aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt
     2821 ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag
     2881 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg
     2941 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac
     3001 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     3061 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     3121 aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat acacagccag
     3181 tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagcattatg tagtctgttt
     3241 tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt ctcgttcagc
     3301 tttcttgtac aaagtggtga tactagttag tctgcgatgc tgtggaccga cgcattgaaa
     3361 tacacaccgt ctttggcgtt cctttttttt tttttgattt gtttttttta ttgagatgtt
     3421 ttgttttgtt tttttcagtt tttattatat cagcagtttg tccgcctgca ttcatgcagt
     3481 gtttggtaat tctttctatt ttttggaatt atggcgataa tattcttgtc tttaaacttc
     3541 ttataaccaa ttgtgcttta gagtttcctg cttagttgct attaacacac tgttaggaac
     3601 gtgaaaccct cgagatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3661 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3721 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3781 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3841 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3901 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3961 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4021 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4081 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4141 cgcgaatgcc cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4201 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4261 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4321 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4381 tctatcgcct tcttgacgag ttcttctgac tcgagtctgc gatgctgtgg accgacgcat
     4441 tgaaatacac accgtctttg gcgttccttt tttttttttt gatttgtttt ttttattgag
     4501 atgttttgtt ttgttttttt cagtttttat tatatcagca gtttgtccgc ctgcattcat
     4561 gcagtgtttg gtaattcttt ctattttttg gaattatggc gataatattc ttgtctttaa
     4621 acttcttata accaattgtg ctttagagtt tcctgcttag ttgctattaa cacactgtta
     4681 ggaacgtgaa accgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
     4741 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4801 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4861 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4921 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4981 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5041 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5101 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5161 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5221 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5281 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5341 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5401 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5461 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5521 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5581 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5641 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5701 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5761 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5821 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5881 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5941 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6001 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6061 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6121 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6181 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6241 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6301 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6361 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6421 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6481 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6541 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6601 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6661 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6721 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6781 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6841 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6901 tgccac
//
