LOCUS       40924_10566        6754 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Gateway cloning destination vector pcGFP1Fpac, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  AUTHORS   Dawson SC, House SA.
  TITLE     Imaging and analysis of the microtubule cytoskeleton in giardia
  JOURNAL   Methods Cell Biol. 97, 307-339 (2010)
  PUBMED    20719278
REFERENCE   2  (bases 1 to 6754)
  AUTHORS   Hagen KD, Hirakawa MP, House SA, Schwartz CL, Pham JK, Cipriano MJ, 
            De La Torre MJ, Sek AC, Du G, Forsythe BM, Dawson SC.
  TITLE     Novel structural components of the ventral disc and lateral crest in
            Giardia intestinalis
  JOURNAL   PLoS Negl Trop Dis 5 (12), e1442 (2011)
  PUBMED    22206034
REFERENCE   3  (bases 1 to 6754)
  AUTHORS   Cipriano MJ, Pham JK, Hagen KD, Dawson SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-MAR-2018) Dept of Microbiology and Molecular Genetics,
            University of California, Davis, One Shields Ave, Davis, CA 95616, 
            USA
REFERENCE   4  (bases 1 to 6754)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6754)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods 
            Cell Biol. 97, 307-339 (2010)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "PLoS Negl 
            Trop Dis"; date: "2011"; volume: "5"; issue: "12"; pages: "e1442"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (12-MAR-2018) Dept of Microbiology and Molecular Genetics, 
            University of California, Davis, One Shields Ave, Davis, CA 95616, 
            USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    893..914
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        929..959
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    967..983
                     /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)."
     primer_bind     991..1007
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1025..1043
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    1067..1191
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1216..1246
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1300..1956
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2301..2603
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2647..2771)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2785..3501
                     /label=EGFP
                     /note="enhanced GFP"
     3'UTR           3514..3638
                     /gene="alpha tubulin"
     gene            3514..3638
                     /gene="alpha tubulin"
                     /label=alpha tubulin
     5'UTR           3717..3919
                     /gene="gamma giardin"
     gene            3717..3919
                     /gene="gamma giardin"
                     /label=gamma giardin
     CDS             3926..4519
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     3'UTR           4529..4646
                     /gene="glutamate dehydrogenase"
     gene            4529..4646
                     /gene="glutamate dehydrogenase"
                     /label=glutamate dehydrogenase
     promoter        complement(4666..4684)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4691..4707)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4848..5303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5635..5739
                     /label=AmpR promoter
     CDS             5740..6597
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      781 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      841 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      901 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      961 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     1021 agctatttag gtgacactat agaatactca agcttggcgc gccatcacaa gtttgtacaa
     1081 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
     1141 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca
     1201 ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt
     1261 taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     1321 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     1381 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1441 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1501 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1561 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1621 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1681 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1741 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1801 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1861 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     1921 ttacaacagt actgcgatga gtggcagggc ggggcgtaaa cgcgtggatc cggcttacta
     1981 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact
     2041 gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga
     2101 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
     2161 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
     2221 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
     2281 cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
     2341 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     2401 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     2461 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     2521 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     2581 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
     2641 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     2701 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg
     2761 tacaaagtgg tgataccggt cctcatggtg agcaagggcg aggagctgtt caccggggtg
     2821 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2881 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2941 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3001 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3061 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3121 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3181 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3241 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3301 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3361 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3421 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3481 ggcatggacg agctgtacaa gtaaagcggc cgcagtgaat tctttccgtc gacctttgca
     3541 tcatcgttgc tcggtcgtct gcctcgttta cgacctgctt gttcctgggc ctcaaatggt
     3601 tagtggtttt ccttcttcca acggcgttgg tccatcctcg gccgcgtgat caaatgcctt
     3661 cgagcgtatg tctgacctgt ttgtgaatga cgatccgtgg ctttactcag attcctgtgt
     3721 ttgtgaataa gaaatccgtg aatttgttgg ggtggaacgg cgaagtggcc tctgctacaa
     3781 taagaactgg agagtgtgtt cagaggcgaa tgcgcaacga actttgtcgt tacacgacgt
     3841 gtgcgcctgt ggtcttcgtg cggtttccgc gcggaatttt gcgccttttc taaaaaacta
     3901 accctaaaac aagaaaatca tgggcaccga gtacaagccc acggtgcgcc tcgccacccg
     3961 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4021 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4081 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4141 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4201 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4261 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4321 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4381 cgccggggtg cctgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4441 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4501 gacccgcaag cccggtgccc tcgagtgagg gctgaagtga atatttacct tttccgggtt
     4561 cgacgggcgt gcttgacgat gcggaagtcg ctgtccgttc caacggctat ctgccccagg
     4621 acccccatcc cggcgccgat cagctggtac cgagctcgaa ttcgccctat agtgagtcgt
     4681 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4741 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4801 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt
     4861 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4921 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4981 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5041 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5101 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     5161 ttgacgggga cagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     5221 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5281 taatgcgccg ctacagggcg cgtcctgatg cggtattttc tccttacgca tctgtgcggt
     5341 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5401 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5461 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5521 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5581 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5701 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5761 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5821 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5881 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5941 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6001 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6061 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6121 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6181 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6241 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6301 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6361 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6421 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6481 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6541 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6601 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6661 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6721 ttttcgttcc actgagcgtc agaccccgta gaaa
//
