LOCUS       40924_30730       10400 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pMF309, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10400)
  AUTHORS   Freitag M, Hickey PC, Raju NB, Selker EU, Read ND.
  TITLE     GFP as a tool to analyze the organization, dynamics and function of 
            nuclei and microtubules in Neurospora crassa
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10400)
  AUTHORS   Freitag M, Selker EU.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-APR-2004) Inst. of Mol. Biology, University of Oregon,
            OR, USA
REFERENCE   3  (bases 1 to 10400)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10400)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-APR-2004) Inst. of Mol. Biology, University of Oregon, OR, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        363..467
                     /label=AmpR promoter
     CDS             468..1325
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1499..2087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2345..2363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2638..2649
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             6934..7650
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(7720..7738)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(join(10399..10400,1..17))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc
       61 gttctaacga caatatgtcc atatggtgca ctctcagtac aatctgctct gatgccgcat
      121 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      181 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
      241 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
      301 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
      361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      481 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      541 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      601 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      661 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      721 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      781 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      841 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      901 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      961 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1021 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1081 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1141 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1201 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1261 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1321 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1381 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1441 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1501 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1561 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1621 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     1681 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1741 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1801 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1861 acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga
     1921 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1981 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2041 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2101 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2161 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2221 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2281 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggttaacc tggcttatcg
     2341 aaattaatac gactcactat agggagaccg gcagatccaa tgcggatgga ttcgctcact
     2401 cggtgccgcg gcggtgcgaa gttcctgccg tggattgggg gattgcacag acggtggagt
     2461 gcttgctaca gctaggtacc cagttgcaca ccaaacggtg cccctgtgcc cctgcgtcta
     2521 ggtgtgtcct gtgagcaact gcgcccgcgc gcggcaccgg gaagctgcgg gaacctcaat
     2581 ggatgattgg cgccagatcc ccggcatcac ctcagcacat gggcagcccg agaaggaatc
     2641 gagggtagaa gatattccag ggtttccgct agcacaactt ccaagcgcct ctaccgctac
     2701 tccacacacc cacgctgaat ggcacaacca ctgtaggcct agcggtccca gctccccgta
     2761 accacgcagc cccagcgacc gaggtcatca tgacactatg acgacaatct ggcgggagac
     2821 ttgggaaaat cgatgacggg cacctacctt gggaacaata gagcaagtcc agtcaacgcc
     2881 ctagggaagg gggaaaaaaa aaaagagccc attctcacaa ggagcaacac tgtgctgcta
     2941 tcatagtaca ctacaagcaa agtcgggatg ttgctagaat ccgtcggcta cgtcggtaga
     3001 tacctcccat aggcagtaag tctaggtatc caatttatag ttccgcgtat ttcttcgtcg
     3061 aacggggcac agcaaagcct aacagccgca gtgcctcgca accaacacca agcagtaatc
     3121 tcatccatcc atacagatcc tcccacaata tctttacatg cgcacttgcg ctgtgttagc
     3181 agtttaggtt ccactaccaa ggctgacacg agcagcactg acagcaaact gctgcttggg
     3241 gccaagaaat tggcagtttg ctgagcacct acatatcagt acacatcaac cctggtttaa
     3301 ttgttagcct ggtactgtta accgtcatga acgaggacgt ggaggaagaa ccacgctaac
     3361 aattgaacca gggttgatgt atacctctct taccatattt gcttgcttcc acgtccctat
     3421 cctcaagcga ttttgttctt gactttttgt atttcaacgc aaaggccatt caggtttctc
     3481 atgttcaacc ctttggatgg cgatgccgtt cgtcccagga cctcgtgacc tttgaaagac
     3541 gagaaatata tcccaaagca acttctgctt atggacaagc aactgcatgg cggcccacaa
     3601 ctcatatctc gcgtaatcat ccaaacgcta ggcccgagta ctctactgtt ctcgagacac
     3661 ctaccataat atgtacccaa tcagacatga caacagtcaa ccaaacatac cctgatccgg
     3721 ccaaaccctg acatccctag atgatggtta ggcacacaaa tggaaaacgg gactcgttgg
     3781 ttaggtggga acgcttgtat tgcagccagc caaccgactc tcctcgtccc tctcgagtcc
     3841 cgttattgcc gtttgaccca gccgcccagc cgccaaacag ctgagggagc caatggtcta
     3901 atacaccgtc caacggcccc agtgatgttt caaacttgat tgacagcgaa cgaaacccct
     3961 gaaacccgtt atagatatgt atagatttga cggggtagct tggcccttaa tacgactcac
     4021 tatagggcga attggagctc caccgcggtg gcggccgcta gaaggagcag tccatctgcg
     4081 tgaatcacga gagaatcagc tactttgaat cgatggatgc agctaccaga agtcactcag
     4141 ttcgttcaaa gccacatcac tgggcacttc cattgggaca ggcattgatc ggacgagacc
     4201 gacttctggc cgctttcaac agccacatta tatccatgtc acggctacgc gtcggccttc
     4261 ggtaacagaa aagcacacag acagcgattg tgacatggat tcgggcaaag cattggtggt
     4321 cgcaccaggt catcctgagt gtgcagtggc tgctattcag atttcatcta actgcgggag
     4381 aggggttcaa aggggcgtga cgtcacagac aacgggtgaa ggacgaagat tgcctcactt
     4441 ctttgctagc aatttgcctg caaagaaggc gcacatgaca agcaaacaac tgggaaacca
     4501 ctattgaata cccacaatgc aaagctcgga aggtacgtct tgattgcagt gtgtcgagtg
     4561 tcaaaaaaga agcaagtgtt catgcaagcc aaaattggca cctcctccac ttctccgagt
     4621 gccccacccg aacctccagg cgagatggcc ggaacatacc atccgcgttg ggattatgac
     4681 gtatctcctt cttcttcaca tgattccatc ccgttgttgc ttgtttgcga gctgtgacgg
     4741 gagatcgtag atgccacttc gggccaggca ggcagtgcag gcagccagga acacaagctt
     4801 ccaacttggt catctcgatt gccgattaag ggaaccaaat gcctatataa gactgtcctc
     4861 ccacctcccc aataccattc ttttcttctt ccatcatcag ccaacaaagc aatcacatct
     4921 tcactacttc aaatcaacac aacactcaaa ccactttcac aacccctcac atcaaccaaa
     4981 tctagaacta gtggatcccc gggaagatgc gtgaaattgt aagtctccct ctgatgccta
     5041 ctgtgccgcg cagcagagca atccctcttt tttttctctt tccctcccaa gagacccctg
     5101 attgctctac cccaccaaac tgcctcgtcc ccttctgtgc ctctacctac tgctgcccga
     5161 ctcattatga tcctgctcgc tttctcgcca tcgcggactg ccatctatcg cgagtattat
     5221 ggacatcgac gctgacacga tttcttcacg tgctataggt tcatctccaa accggccaat
     5281 gcgtaagtcg atgatggctt ccacccccgt cgcgatccaa caacaacagc aggactaaca
     5341 caacagccac gatcagggta accaaatcgg tgctgctttc tggtacgtcg tcatcatcgc
     5401 gaacaaccac acctttgagc gagggagccc gacgctgaca gaataaacag gcagactatc
     5461 tccggcgagc acggcctcga tgcctccggt gtgtacgtaa cctcgtgccc aataaccata
     5521 tctggtgatg gcctcgaaag ctcaccgccc actacaggta caatggcacc tctgagctcc
     5581 agctcgagcg catgaacgtc tacttcaacg aggtgagcaa caaccggtct cgacgatctc
     5641 cccctcctcg agaattcgct cgctaactag cttcgcgctt gacaggcttc cggcaacaag
     5701 tatgtccctc gtgccgtcct cgtcgatctc gagcccggta ccatggacgc cgttcgcgcc
     5761 ggtcccttcg gccagctctt ccgccccgat aacttcgtct tcggccagtc cggtgctggc
     5821 aacaactggg ccaagggtca ctacactgag ggtgctgagc ttgttgacca ggttctcgat
     5881 gtcgtccgtc gcgaggctga gggctgcgac tgcctccagg gcttccagat cacccactcc
     5941 ctcggtggtg gtaccggtgc cggtatgggt accctcctta tctccaagat tcgtgaggag
     6001 ttccccgacc gcatgatggc tacctactcc gtcgtgccct cccccaaggt ctccgatacc
     6061 gttgtcgagc cctacaacgc caccctctcc gtccatcagc tcgttgagaa ctccgacgag
     6121 accttctgca ttgacaacga ggcgctttac gacatttgca tgaggaccct caagctctcc
     6181 aacccctctt acggcgatct taaccacctc gtctccgccg tcatgtccgg tgtcaccgtc
     6241 tccctccgtt tccccggcca gctgaactcc gatctccgca agctcgccgt caacatggtt
     6301 cccttccccc gtctccactt cttcatggtc gggttcgctc cccttaccag ccgcggcgcc
     6361 caccatttcc gtgccgtctc cgtgcccgag ttgacccagc agatgttcga ccccaagaac
     6421 atgatggctg cttctgactt ccgcaacggt cgttacctca cctgctctgc catcttgtaa
     6481 gttgctgatc atggaatgct gcttttgacc actaatactg acgaatcaaa cagccgtggc
     6541 aaggtctcca tgaaggaggt tgaggaccag atgcgcaacg ttcagaacaa gaactcttcc
     6601 tacttcgtcg agtggatccc caacaacgtc cagactgccc tctgctctat ccctccccgc
     6661 ggtctcaaga tgtcctccac cttcgtcggt aactccaccg ccatccagga gctcttcaag
     6721 cgtatcggcg agcagttcac tgccatgttc aagcgcaagg ctttcttgca ttggtacact
     6781 ggtgagggta tggacgagat ggagttcact gaggctgagt ccaacatgaa cgatctcgtc
     6841 tccgagtacc agcagtacca ggatgctggt gttgacgagg aggaggagga gtacgaggag
     6901 gaggcccccc ttgagggcga ggagttaatt aacatggtga gcaagggcga ggagctgttc
     6961 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     7021 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     7081 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcac ctacggcgtg
     7141 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     7201 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     7261 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     7321 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     7381 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     7441 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     7501 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     7561 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     7621 atcactctcg gcatggacga gctgtacaag taagaattcg atatcaagct tatcgatacc
     7681 gtcgacctcg agggggggcc cggtacccag cttttgttcc ctttagtgag ggttaattag
     7741 ggcgtggacg gctaatgggg tctgaatgct aaaagacacc atttcccaca ctccctctga
     7801 ttttttttat ggtttttttt tatcttcatt tttcgccgtc ttttattcgg ttgtttggga
     7861 ggttcacatg caacgtgcaa gactgcaagt tttttgagat gggcgcttca tgcaatgcag
     7921 catcactgca gggacattgt gacatccacg tttcgatcgt ctgtgcttcc cccgttcatg
     7981 tacgcatgat ctgaaaaaaa aaaaaaaaaa aaaggttgat gtaataatga aaataaacac
     8041 ccagggacga gcacctcatc tgctggaata gggagaaaaa ctcaagggcc gtttgggtct
     8101 ccacacttac caacagattc atgtactgat cttcagcctg tacagaacag tacaatgtcc
     8161 aaaactcgtt atcattactc tatgtgccca aaatgaccca gatatacatc aacccactac
     8221 atccttttgt tcttgttttt tttttttttt tttgctactt cgaaaagaat ttccgtttag
     8281 ttgctcttgc tcatgtgctc aagacggatg ctgaccgccc ttctgtgagc ctcgagctcc
     8341 tcaaccttag ccaactgcat gacagcctgg ccgacgtttt tgagaccctc ggcagtcaag
     8401 ttggagctgg taatgtgctt gacgaacgag gcgaaattga cgccagagta ctgcttgcca
     8461 aagccatagg tagctatgtg tcttgacgtt agcagaagaa tctgtgaagc ggggctattt
     8521 atgatatgta acttacgcag cgagtggtta acaccagcag agtaatcgcc aacggactca
     8581 ggagtccagg cgccaatgaa gacactaccg gcgttcatga caagatcgac agccttctcg
     8641 gcctccttga tctggaggat caagtgctca ggagcgtact tgttgctgag ctccatggcc
     8701 tcctcgacgg tcttgacctg cacggtgatc gagtgggcga tggagccacg gacaatctgg
     8761 acgcgaggaa gctccgtagc ctgacggtga acctcgtcct caatagcctg aagatgctcc
     8821 tcgtcgaggt caatagcgat caggatgacc tgactgtcaa cgccgtgctc agcctgggac
     8881 aggagatccg aggcaacgaa cgcggggttg gcgtccttgt cagcgatgac cagcacctcg
     8941 gacgggccag cgggcatgtc aataccaacg gcagcgttgg tgtcgttgct gacgaacatc
     9001 ttggcagcag tgacgaactg gttaccgggg ccgagaatct tgtcgacctt ggtgatgctc
     9061 tcggtgccgt aggccatggc agctacggcc tgggcaccgc cggcaagcac gatggactcg
     9121 gccccaacct tgtgagcgac gtggacaatc tcgggagtga tggttccgtc ggcgcgggga
     9181 ggagaggcga acacaatctt gttgcagccg gcgaccatgg cgggaacacc cagcataagg
     9241 gcagtgctgg ggagaacggc ggtaccgccg gggatgtagc agccgacggc ctcgatggga
     9301 cgagagaaac ggctgcagac aacaccgggc atggtctcga cctggagggg cttctcctcc
     9361 ttctgggcgg cgtggaactt gcggatgttc tcgaaggaca cgtcgatggc agcaatggtc
     9421 tcctcaggga gctgcataag ctccttgggg aagggcgcct tcaggacggg gctagtaaga
     9481 gaggtagcct tctcgaactt gtgagtgtac gacagaacag ccttgtcgcc gttcttgcgg
     9541 acgtcctcga tgatggggac aatgatcttg tagatggcat cggacgactt ttgcgcagga
     9601 cgcttgagag cagcatcgag ctcctcggta gagaccttgg aggcgtcgaa acgtctcatg
     9661 gtgatcttct caggggtggt ctcctgggcg gcctccttgg tgacaggggc cgaagtggca
     9721 gcgagagcgg acgccgcagg cttgatgccc tccttctcag cccacttacc cttagcatct
     9781 ccagtcctgc gcttgacctt ccagctcttg gcgtcaaggc tcctttcgat atcggcaaga
     9841 gtaacgccgg cagcaacggc cctggtaaga gcaaagtaga agagatcggc agcctcaaag
     9901 gcgatttcct ggggggtctg agcggtgcag agctcctcag cctcctccat gatcttggcc
     9961 cggactagct tctcatcgga gaagagacgg gcagtgtagg agccctcggg ggcagactgt
    10021 ttcctcgaaa tcaaagtctg ctcgagcttg ggaaggcctt tgagctgacc aaagcagccg
    10081 gactgatcga ggtggcagaa acgacccttc tgcttcacga caaacttgag agcatcgtta
    10141 tcgcagtcaa gcgagatgcg gacgagctcc tgagtgtctc cggaagtagc acccttgtac
    10201 cagagaccgc gcttccggct ctgatagaca ccagtctgtg tcctgagggc ctcgttcaca
    10261 ctctcggcac tgctgtagac cagacccaga gcagtgtcgt gctcatcaac gacaacggtg
    10321 gggagaagac catcgggacg gtcagacttc cagacgctcg agaggatggt ggagatggca
    10381 agcttcagct gctcgagttc
//
