LOCUS       40924_24588        9649 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector phis-5GFP5', complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9649)
  AUTHORS   Bowring FJ, Yeadon PJ, Catcheside DE.
  TITLE     Use of fluorescent protein to analyse recombination at three loci in
            Neurospora crassa
  JOURNAL   Fungal Genet. Biol. 49 (8), 619-625 (2012)
  PUBMED    22691725
REFERENCE   2  (bases 1 to 9649)
  AUTHORS   Bowring FJ, Yeadon J, Catcheside DEA.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUN-2012) School of Biological Sciences, Flinders 
            University, Sturt Road, Bedford Park, SA 5162, Australia
REFERENCE   3  (bases 1 to 9649)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9649)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fungal 
            Genet. Biol."; date: "2012"; volume: "49"; issue: "8"; pages: 
            "619-625"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-JUN-2012) School of Biological Sciences, Flinders University, 
            Sturt Road, Bedford Park, SA 5162, Australia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9649
                     /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"
     misc_feature    336..1578
                     /function="targeting to the his-5 locus"
                     /label=truncated his-5 gene
                     /note="truncated his-5 gene"
     regulatory      1640..2561
                     /gene="ccg-1"
                     /label=Neurospora crassa ccg-1 (grg-1) promoter
                     /note="Neurospora crassa ccg-1 (grg-1) promoter"
                     /regulatory_class="promoter"
     gene            1640..2561
                     /gene="ccg-1"
                     /label=ccg-1
     primer_bind     2562..2578
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3784..4482
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    4494..6013
                     /function="targeting to the his-5 locus"
                     /label=3' flank ofNeurospora crassa his-5 gene
                     /note="3' flank ofNeurospora crassa his-5 gene"
     promoter        complement(6083..6101)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6108..6124)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6265..6693
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             7037..7828
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             7849..8706
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8880..9468
                     /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 ggcttcaaac atcctcgcct tggaacccta
      361 tcgctgcgcg cgcgagtgag ctcaccccca aaacttctct gtctatgacc tctcatgaag
      421 tacatcgact aacacacgta tttcagtgac tacaaggacg atggcaccaa catcctccta
      481 gacgccaatg agaacgccta tgggcctccc gttccccaga acctggctac cggctccggc
      541 tccagtggtc ccgagcttga cctcctcggc ctgcaccgct accccgaccc tcaccagcgg
      601 gacctcaagc agctcctctg ctcgctgcgc acgacccacg cgcacacctc caagcttctg
      661 acgcccgaga acctctgcgt aggcgtcggc tctgacgagg ccatcgacgc tctcttgcgg
      721 tgcttctgcg ttcccggccg cgaccgcatc ctcgtctgcc cgcccaccta cggcatgtac
      781 tccgtttccg ctcaggtgaa cgacgtcgga ctcgtcaaag tcccgctgct ccccgctccc
      841 tccttcgcca tcgacgtccc gcgcgtcctc gagacgctgt cgaccgaacc caacatcaag
      901 atcatctacc tctgctcgcc gggtaacccc acgggatccc ttctcgccaa ggaagacatc
      961 caaaaggttc tcgaacaccc gacctggaac ggcgtcgtgg tcgtcgacga ggcctacatt
     1021 gactttgcgc ctgagtcggc ctcgctggcc gagtgggtca acgagtggcc caacctggtc
     1081 gtcatgcaga ctttgtccaa ggcgttcggc ctggccggca tccgtctggg agcggccttt
     1141 acaagtcccc ctatcgccag gctgctgaac agcctcaagg ctccctacaa catttcttcg
     1201 ccgaccagcg cgctggccag ctacgcgctg ggaccggacg gactaaaggt gatgcgtgcg
     1261 aacaaggaga ggctgattaa gcagcgggag aggctggtgg cggagctgcc gcagattccg
     1321 ggcgtgggca agttggcggg cggtgagagc agcaactttt tgttgtacga gatgctggac
     1381 aaggagggaa agccgagcaa cgagacggcg ctgaaggtgt atcagaagtt ggccgagacg
     1441 aagggagtgg tggtccggtt cagaggcaag gagcacgggt gcgaggggtg cttgaggatt
     1501 acggtgggta cggaagagga ggttacgagg tgcctgggag cgttgaggga ggcgttgaag
     1561 gaggttaggg gggagtaagt gtggtcatca ggtgtcgagg gagatgttcc tttcgaacgc
     1621 aatcgtcggc gcgggccgct agaaggagca gtccatctgc gtgaatcacg agagaatcag
     1681 ctactttgaa tcgatggatg cagctaccag aagtcactca gttcgttcaa agccacatca
     1741 ctgggcactt ccattgggac aggcattgat cggacgagac cgacttctgg ccgctttcaa
     1801 cagccacatt atatccatgt cacggctacg cgtcggcctt cggtaacaga aaagcacaca
     1861 gacagcgatt gtgacatgga ttcgggcaaa gcattggtgg tcgcaccagg tcatcctgag
     1921 tgtgcagtgg ctgctattca gatttcatct aactgcggga gaggggttca aaggggcgtg
     1981 acgtcacaga caacgggtga aggacgaaga ttgcctcact tctttgctag caatttgcct
     2041 gcaaagaagg cgcacatgac aagcaaacaa ctgggaaacc actattgaat acccacaatg
     2101 caaagctcgg aaggtacgtc ttgattgcag tgtgtcgagt gtcaaaaaag aagcaagtgt
     2161 tcatgcaagc caaaattggc acctcctcca cttctccgag tgccccaccc gaacctccag
     2221 gcgagatggc cggaacatac catccgcgtt gggattatga cgtatctcct tcttcttcac
     2281 atgattccat cccgttgttg cttgtttgcg agctgtgacg ggagatcgta gatgccactt
     2341 cgggccaggc aggcagtgca ggcagccagg aacacaagct tccaacttgg tcatctcgat
     2401 tgccgattaa gggaaccaaa tgcctatata agactgtcct cccacctccc caataccatt
     2461 cttttcttct tccatcatca gccaacaaag caatcacatc ttcactactt caaatcaaca
     2521 caacactcaa accactttca caacccctca catcaaccaa atctagaact agtggatcca
     2581 ccatgcctcc caagaagacc gagaccaagg ccgccgatgc ctctgccgcc gccgctcctg
     2641 ctcccgccgc tgctcccacc tctgctccca agaccaagag ccccagcacc catgcctctt
     2701 accttgtaag ctttacttaa tccccaagag aaacgcatct tttgctaaca ttcaaacagg
     2761 acatgatcac ggacgctatt gttgcggtat ttaaaccccc tcattatctt tgttgtggcg
     2821 tgttttggtt gtgggcgctt ctctggtcgg tgagcaaaat gagacaattg ctctcatgat
     2881 gcacgcaacc aagagtcacc cacaccactc accacccttt tactgtccct cagtcgtgct
     2941 gattccttct ttttgtcatg ctacagctca aggaccgcgc tggatctagg tatgtgccat
     3001 ttttctgctt acttttctca ttttttcacc agggctttgc caagccacac cgcaaacctt
     3061 gcgggccctt cctacggtcg caccgatgac cactttcatt cttatttcca tgacgcgatg
     3121 aaggatcaca acatgttgct aacgcacacc acagccgtca agctctcaag aagtatgtcc
     3181 gtgccaacaa cacgctcggc aacgtcaccg acaacatgtt cgactcgctc ttcaacaagg
     3241 ccttgaagaa cggtgtcgac aagggtgtct ttgagcagcc caagggtatg tcaccaccca
     3301 catgacgcat catctccttg tattactctt tgcatgctca cacacgatat aggtccttcc
     3361 ggtggcacca agcttgccaa gaaggtcgct aagcctgctc ccaagaaggc tgctcccaag
     3421 aaggagacca aggagaagaa gcccgctgcc gctaagaagg agggtgccgc caagaaggag
     3481 accaaggaga agaaggctcc tgctgccaag aaggcggctg cccccaagaa ggctgctgct
     3541 cccaagaagg aggtcaagga gaagaaggcg gctgccccca agaagaaggc cgctgctcct
     3601 gccgttgccg acaaggagac cgtcctcacc aagaccaagt ctggccgtgt cgccaagagc
     3661 accgctaagc ctgctgccgc caagaaggcc gctgctccca agaaggctgc cgccagcaag
     3721 aaggccgaga aggccgagcc cgctgccgag aaggcattaa ttaacatggt gagcaagggc
     3781 taggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3841 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3901 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     3961 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     4021 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     4081 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     4141 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     4201 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     4261 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     4321 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4381 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4441 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagaatt cgcgccccgt
     4501 attcggtgct ggcatgtaac caactgggga aggagtaggt tcaggacaaa agaacacaac
     4561 aatgcaatac actagtttca ccctgcttta ccatactccg ttacaacctg gactggaaag
     4621 tatgcgaaag tgcattcgcg ttcaccttgc tacctagtta acatgctaaa tatacaaagt
     4681 caacaccacg ctcaacttgc gaccaaaaaa gtaaacacta taaaagtgaa cagcaggggt
     4741 cagctcagca tattggactt tgcggcaaaa aatgcaaaag atcaggtcgt acccggaatc
     4801 gaaccggggt tgccggaatg ttctgaagaa atcagaatcc ggagtgataa ccgctacact
     4861 atacaacctt tgatggtcta aacgtactcc acattgcatc tatgagcact ctgtgtactg
     4921 gtcacacatc gtgcaaagcg ctgtcaaaca atcgacgttt cggactttta agacattgca
     4981 caaaagttcg ttacaaacaa tatgacacgt acggagtttg cttaggtaac ttgatagaca
     5041 ataacccaat actccggaaa cccaggaaaa tgaaatgtcg cgactccacc ctcgtttgtg
     5101 ctcgttaagt tcgcggaagt catcatccac tatcaggttg cacaccatcc catcttcacc
     5161 cttgtcttcg ccaattttcc acatgcgttc tttgccacga gggtatgata aactgaatat
     5221 tagaaagaaa ccactcaatc caccaactca tccattgcta acgtctattt gtctcctcca
     5281 ccaactcgac ttggctcttt gctggtcgcg agctggcaac taaacaacaa cttctgttga
     5341 agatcagcca ccgcgccata ccaacggggt atcgtgatca tggaatcccg gactttgccg
     5401 gccatggtgt attcaggtaa agtgtgaaac cgaccacagc atgatttttg ccgtagaagg
     5461 tttaaccaat gaagctcgct taaatcagca gaggctgctg ctcaccgctc ttgacagtga
     5521 cgggaacgac aggctcggtc tggatgtcgg cctggggggc gggaggaaag tcggcgtcgc
     5581 ggagctcgga cttgttggca tgaccgttgg tgtgctcctc accgtggaga gaggcaggag
     5641 tagccttgct gaccagcgtc tgggccttgg cggtatactc gccaacaaac tgcttggtca
     5701 actgggtagc cttctcgctg ttctgctgag ccgcggtctt gagctgagca gtctgggcgc
     5761 cgatgatctc gctagcacgg tgaagctggg catcgataaa ctccttgttg gtcttgtaga
     5821 cgaggggccc gaggaaggca agagtggtgg caatcagggc aagaccccag tagggaacga
     5881 tcttgacgag ataataggcg aagaaagcgg cgatggcaac gaggccggac acgtagacat
     5941 tttcagcgaa gagaatacgc tgagcctcaa tgacgaagaa gttgacgagc tcatgaacat
     6001 cgccaatagc gctaagccga attctgcaga tatccatcac actggcggcc gctcgagcat
     6061 gcatctagag ggcccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
     6121 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     6181 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6241 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     6301 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     6361 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     6421 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6481 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6541 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6601 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6661 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aattcagggc gcaagggctg
     6721 ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     6781 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc
     6841 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc
     6901 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
     6961 ggctttcttg ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg
     7021 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     7081 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     7141 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     7201 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     7261 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     7321 cgaagtgccg gggcaggatc tcctgtcatc ccaccttgct cctgccgaga aagtatccat
     7381 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     7441 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     7501 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     7561 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     7621 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     7681 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     7741 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     7801 cttctatcgc cttcttgacg agttcttctg aattgaaaaa ggaagagtat gagtattcaa
     7861 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7921 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7981 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     8041 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     8101 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     8161 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     8221 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     8281 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     8341 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     8401 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     8461 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     8521 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     8581 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8641 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8701 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8761 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8821 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8881 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8941 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     9001 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     9061 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     9121 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     9181 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     9241 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     9301 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     9361 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     9421 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     9481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     9541 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     9601 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
