LOCUS       40924_43168        6695 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Neurospora cloning vector pTH1067.9, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6695)
  AUTHORS   Hammond TM, Xiao H, Rehard DG, Boone EC, Perdue TD, Pukkila PJ, Shiu
            PK.
  TITLE     Fluorescent and bimolecular-fluorescent protein tagging of genes at 
            their native loci in Neurospora crassa using specialized 
            double-joint PCR plasmids
  JOURNAL   Fungal Genet. Biol. 48 (9), 866-873 (2011)
  PUBMED    21664475
REFERENCE   2  (bases 1 to 6695)
  AUTHORS   Hammond TM, Shiu PKT.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2011) Biological Sciences, University of 
            Missouri-Columbia, 103 Tucker Hall, Columbia, MO 65211, USA
REFERENCE   3  (bases 1 to 6695)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6695)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fungal 
            Genet. Biol."; date: "2011"; volume: "48"; issue: "9"; pages: 
            "866-873"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-MAR-2011) Biological Sciences, University of Missouri-Columbia, 
            103 Tucker Hall, Columbia, MO 65211, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        342..696
                     /label=trpC promoter
                     /note="promoter for Aspergillus nidulans trpC"
     CDS             701..1723
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     regulatory      complement(1755..2314)
                     /label=trpC
                     /note="trpC"
                     /regulatory_class="terminator"
     CDS             complement(2318..3034)
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    complement(3035..3058)
                     /label=4X(GlyAla)-linker
                     /note="4X(GlyAla)-linker"
     promoter        complement(3129..3147)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3154..3170)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3311..3739
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             4083..4874
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4895..5752
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5926..6514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttgtta actgatattg aaggagcatt
      361 ttttgggctt ggctggagct agtggaggtc aacaatgaat gcctattttg gtttagtcgt
      421 ccaggcggtg agcacaaaat ttgtgtcgtt tgacaagatg gttcatttag gcaactggtc
      481 agatcagccc cacttgtagc agtagcggcg gcgctcgaag tgtgactctt attagcagac
      541 aggaacgagg acattattat catctgctgc ttggtgcacg ataacttggt gcgtttgtca
      601 agcaaggtaa gtggacgacc cggtcatacc ttcttaagtt cgcccttcct ccctttattt
      661 cagattcaat ctgacttacc tattctaccc aagcatccaa atgaaaaagc ctgaactcac
      721 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
      781 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
      841 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
      901 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
      961 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     1021 actgcccgct gttctccagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     1081 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     1141 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     1201 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     1261 ctgccccgaa gtccggcacc tcgtgcatgc ggatttcggc tccaacaatg tcctgacgga
     1321 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     1381 cgaggtcgcc aacatcctct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     1441 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cgcctccggg cgtatatgct
     1501 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     1561 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     1621 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     1681 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagagta gatgccgacc
     1741 gggaaccagt taacctgtgc attctgggta aacgactcat aggagagttg taaaaaagtt
     1801 tcggccggcg tattgggtgt tacggagcat tcactaggca accatggtta ctattgtata
     1861 cccatcttag taggaatgat tttcgaggtt tatacctacg atgaatgtgt gtcctgtagg
     1921 cttgagagtt caaggaagaa acagtgcaat tatctttgcg aacccagggg ctggtgacgg
     1981 aattttcata gtcaagctat cagagtaaag aagaggagca tgtcaaagta caattagaga
     2041 caaatatata gtcgcgtgga gccaagagcg gattcctcag tctcgtaggt ctcttgacga
     2101 ccgttgatct gcttgatctc gtctcccgaa aatgaaaata gactctgcta agctattctt
     2161 ctgcttcgcc ggagcctgaa gggcgtacta gggttgcgag gtccaatgca ttaatgcatt
     2221 gcagatgagc tgtatctgga agaggtaaac ccgaaacgcg ttttattctt gttgacatgg
     2281 agctattaaa tcactagaag gcactctttg ctgcttactt gtacagctcg tccatgccga
     2341 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
     2401 ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc
     2461 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
     2521 tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat
     2581 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     2641 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     2701 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     2761 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga
     2821 agcactgcac gccgtaggtg aaggtggtca cgagggtggg ccagggcacg ggcagcttgc
     2881 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     2941 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     3001 ccccggtgaa cagctcctcg cccttgctca ccatggctcc agcgcctgca ccagctccaa
     3061 gggcgaattc tgcagatatc catcacactg gcggccgctc gagcatgcat ctagagggcc
     3121 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
     3181 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3241 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3301 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3361 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3421 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3481 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3541 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3601 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3661 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3721 caaaaattta acgcgaattt taacaaaatt cagggcgcaa gggctgctaa aggaagcgga
     3781 acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     3841 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
     3901 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     3961 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     4021 caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc
     4081 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4141 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4201 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4261 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4321 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4381 aggatctcct gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4441 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4501 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4561 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4621 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4681 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4741 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4801 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4861 ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4921 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4981 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5041 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5101 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5161 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5221 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5281 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5341 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5401 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5461 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5521 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5581 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5641 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5701 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5761 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5821 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5881 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5941 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6001 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6061 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6121 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6181 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6241 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6301 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6361 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6421 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6481 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6541 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6601 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6661 cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
