LOCUS       40924_20041        8790 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pFGWDR196, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8790)
  AUTHORS   Li X, Romanowsky SM, Harper JF, Cheung AY, Sze H.
  TITLE     Thapsigargin-Sensitive Ca2+/Mn2+ Pump AtECA3 Supports Pollen Tip 
            Growth and Promotes Male Fertility
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8790)
  AUTHORS   Li X, Sze H.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2007) Cell Biology and Molecular Genetics, 
            University of Maryland, H J Patterson Hall Rm 3232, College Park, MD
            20742, USA
REFERENCE   3  (bases 1 to 8790)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8790)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-MAR-2007) Cell Biology and Molecular Genetics, University of 
            Maryland, H J Patterson Hall Rm 3232, College Park, MD 20742, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..722
                     /label=EGFP
                     /note="enhanced GFP"
     protein_bind    729..853
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        878..908
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             962..1618
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1963..2265
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2309..2433)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2470..2488)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2636..2823
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2835..2851)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3287..4633)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4747..5547)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5548..5763)
                     /label=URA3 promoter
     promoter        5874..5978
                     /label=AmpR promoter
     CDS             5979..6836
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7010..7598
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7886..7907
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7922..7952
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7960..7976
                     /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     7984..8000
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      8012..8785
                     /note="PMA1 promoter; derived from yeast; causes
                     constitutive overexpression in yeast"
                     /regulatory_class="promoter"
ORIGIN
        1 ctagtatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
       61 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      121 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      181 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      241 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      301 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      361 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      421 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
      481 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
      541 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
      601 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
      661 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
      721 aggatatcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
      781 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
      841 tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg cttccggctc
      901 gtataatgtg tggattttga gttaggatcc ggcgagattt tcaggagcta aggaagctaa
      961 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1021 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1081 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     1141 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     1201 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     1261 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     1321 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     1381 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     1441 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     1501 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtctgtg atggcttcca
     1561 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     1621 aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     1681 ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtgtgct
     1741 atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat
     1801 atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc
     1861 gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg
     1921 aaatgaacgg ctcttttgct gacgagaaca gggactggtg aaatgcagtt taaggtttac
     1981 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
     2041 acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
     2101 tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
     2161 gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc
     2221 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc
     2281 cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagta
     2341 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta
     2401 cgtttctcgt tcagctttct tgtacaaagt ggtgatatcc ctcgaggggg ggcccggtac
     2461 ccaattcgcc ctatagtgag tcgtattacg cgcggatcca gctttggact tcttcgccag
     2521 aggtttggtc aagtctccaa tcaaggttgt cggcttgtct accttgccag aaatttacga
     2581 aaagatggaa aagggtcaaa tcgttggtag atacgttgtt gacacttcta aataagcgaa
     2641 tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa
     2701 attttaaagt gactcttagg ttttaaaacg araattctta ttcttgagta actctttcct
     2761 gtaggtcagg ttgctttctc aggtatagca tgaggtcgct cttattgacc acacctctac
     2821 cggcatgcca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2881 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2941 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     3001 cggtattttc tccttacgca tctgtgcggt atttcacacc gcataatcgg atcgtacttg
     3061 ttacccatca ttgaattttg aacatccgaa cctgggagtt ttccctgaaa cagatagtat
     3121 atttgaacct gtataataat atatagtcta gcgctttacg gaagacaatg tatgtatttc
     3181 ggttcctgga gaaactattg catctattgc ataggtaatc ttgcacgtcg catccccggt
     3241 tcattttctg cgtttccatc ttgcacttca atagcatatc tttgttaacg aagcatctgt
     3301 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     3361 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     3421 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3481 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     3541 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     3601 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     3661 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     3721 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     3781 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     3841 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     3901 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     3961 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     4021 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     4081 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     4141 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     4201 ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     4261 actttctagc tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga
     4321 gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta
     4381 tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg
     4441 cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct
     4501 gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct
     4561 gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct
     4621 ttgatattgg atcgatccga tgataagctg tcaaacatga gaattgggta ataactgata
     4681 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
     4741 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
     4801 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
     4861 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
     4921 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
     4981 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
     5041 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
     5101 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
     5161 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
     5221 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
     5281 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
     5341 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     5401 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     5461 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     5521 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg tttttgttct
     5581 gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat
     5641 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     5701 atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     5761 gaaaagctaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5821 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5881 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5941 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6001 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6061 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6121 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6181 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6241 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6301 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6361 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6421 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6481 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6541 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6601 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6661 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6721 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6781 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6841 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6901 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6961 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7021 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7081 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7141 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7201 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7261 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7321 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7381 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7441 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7501 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7561 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7621 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7681 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7741 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7801 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     7861 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     7921 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     7981 acacaggaaa cagctatgac catgattacg ccaagcttcc tgaaacggag aaacataaac
     8041 aggcattgct gggatcaccc atacatcact ctgttttgcc tgaccttttc cggtaatttg
     8101 aaaacaaacc cggtctcgaa gcggagatcc ggcgataatt accgcagaaa taaacccata
     8161 cacgagacgt agaaccagcc gcacatggcc ggagaaactc ctgcgagaat ttcgtaaact
     8221 cgcgcgcatt gcatctgtat ttcctaatgc ggcacttcca ggcctcgatc gagaccgttt
     8281 atccattgct tttttgttgt ctttttccct cgttcacaga aagtctgaag aagctatagt
     8341 agaactatga gctttttttg tttctgtttt cctttttttt ttttttacct ctgtggaaat
     8401 tgttactctc acactcttta gttcgtttgt ttgttttgtt tattccaatt atgaccggtg
     8461 acgaaacgtg gtcgatggtg ggtaccgctt atgctcccct ccattagttt cgattatata
     8521 aaaaggccaa atattgtatt attttcaaat gtcctatcat tatcgtctaa catctaattt
     8581 ctcttaaatt ttttctcttt ctttcctata acaccaatag tgaaaatctt tttttcttct
     8641 atatctacaa aaactttttt tttctatcaa cctcgttgat aaattttttc tttaacaatc
     8701 gttaataatt aattaattgg aaaataacca ttttttctct cttttataca cacattcaaa
     8761 agaaagaaaa aaaatatacc ccagcctcga
//
