LOCUS       40924_42719        7820 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Protein fusion vector pTcCFPH-CO 1.1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7820)
  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 7820)
  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 7820)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7820)
  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..7820
                     /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"
     protein_bind    1569..1688
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1718..1748
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1802..2458
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2803..3105
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3149..3273)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3281..3997
                     /label=CFP
                     /note="cyan variant of GFP"
     CDS             4298..5320
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     promoter        complement(5632..5650)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5671..5687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5695..5711)
                     /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(5719..5749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5764..5785)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6073..6661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6835..7692)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7693..7797)
                     /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 cgtgaaacct
     1561 ctagaatcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
     1621 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     1681 tccagtcata ttggcggccg cattaggcac cccaggcttt acactttatg cttccggctc
     1741 gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa
     1801 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1861 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1921 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     1981 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     2041 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     2101 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2161 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2221 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2281 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2341 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2401 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2461 aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     2521 ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct
     2581 atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat
     2641 atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc
     2701 gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg
     2761 aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac
     2821 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
     2881 acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
     2941 tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
     3001 gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc
     3061 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc
     3121 cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagca
     3181 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta
     3241 cgtttctcgt tcagctttct tgtacaaagt ggtgatacta gtaatgagta aaggagaaga
     3301 acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa
     3361 attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt
     3421 tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta ctttgacttg
     3481 gggtgttcaa tgcttttcaa gatacccaga tcatatgaaa cggcatgact ttttcaagag
     3541 tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta
     3601 caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa
     3661 aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat acaactatat
     3721 ttcacacaat gtatacatca ctgcagacaa acaaaagaat ggaatcaaag ctcatttcaa
     3781 aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac
     3841 tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc
     3901 cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc
     3961 tgctgggatt acacatggca tggatgaact atacaaatga ggcgcgccat tgatctgcga
     4021 tgctgtggac cgacgcattg aaatacacac cgtctttggc gttccttttt ttttttttga
     4081 tttgtttttt ttattgagat gttttgtttt gtttttttca gtttttatta tatcagcagt
     4141 ttgtccgcct gcattcatgc agtgtttggt aattctttct attttttgga attatggcga
     4201 taatattctt gtctttaaac ttcttataac caattgtgct ttagagtttc ctgcttagtt
     4261 gctattaaca cactgttagg aacgtgaaac caccggtatg aaaaagcctg aactcaccgc
     4321 gacgtctgtc gagaagtttc tgatcggaaa gttcgacagc gtctccgacc tgatgcagct
     4381 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     4441 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     4501 atcggccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     4561 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     4621 gcccgctgtt cttcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
     4681 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     4741 tgatttcatt tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     4801 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     4861 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     4921 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     4981 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     5041 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
     5101 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     5161 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     5221 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     5281 tggaaaccga cgcctcagca ctcgtccgag ggcaaaggaa tagctcgagt ctgcgatgct
     5341 gtggaccgac gcattgaaat acacaccgtc tttggcgttc cttttttttt ttttgatttg
     5401 ttttttttat tgagatgttt tgttttgttt ttttcagttt ttattatatc agcagtttgt
     5461 ccgcctgcat tcatgcagtg tttggtaatt ctttctattt tttggaatta tggcgataat
     5521 attcttgtct ttaaacttct tataaccaat tgtgctttag agtttcctgc ttagttgcta
     5581 ttaacacact gttaggaacg tgaaaccggg cccggtaccc agcttttgtt ccctttagtg
     5641 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5701 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5761 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5821 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5881 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5941 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6001 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6061 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6121 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6181 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6241 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6301 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6361 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6421 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6481 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6541 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6601 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6661 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6721 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6781 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6841 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6901 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6961 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7021 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7081 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7141 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7201 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7261 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7321 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7381 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7441 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7501 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7561 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7621 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7681 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7741 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7801 atttccccga aaagtgccac
//
