LOCUS       40924_48068        8497 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pYESDR196, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8497)
  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  JOURNAL   Unpublished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 8497)
  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 8497)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8497)
  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            "Unpublished"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..8497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..26
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    44..168
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        220..250
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             304..960
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1305..1607
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1651..1775)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1798..1839
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             1849..1866
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      1900..2147
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2180..2198)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2346..2533
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2545..2561)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2997..4343)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4457..5257)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5258..5473)
                     /label=URA3 promoter
     promoter        5584..5688
                     /label=AmpR promoter
     CDS             5689..6546
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6720..7308
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7596..7617
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7632..7662
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7670..7686
                     /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     7694..7710
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      7722..8495
                     /note="PMA1 promoter; derived from yeast; causes
                     constitutive overexpression in yeast"
                     /regulatory_class="promoter"
ORIGIN
        1 cgactagtaa tacgactcac tatagggaat attaagctat caaacaagtt tgtacaaaaa
       61 agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa
      121 aaaacagact acataatact gtaaaacaca acatatccag tcactatggc ggccgcgggt
      181 gatgctgcca acttagcggc cgcattaggc accccaggct ttacacttta tgcttccggc
      241 tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct
      301 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
      361 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
      421 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
      481 attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac
      541 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
      601 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
      661 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
      721 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
      781 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
      841 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtctg tgatggcttc
      901 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
      961 taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat
     1021 ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtgtg
     1081 ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat
     1141 atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct
     1201 gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat
     1261 tgaaatgaac ggctcttttg ctgacgagaa cagggactgg tgaaatgcag tttaaggttt
     1321 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     1381 acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     1441 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     1501 ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc
     1561 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct
     1621 ccgttataca cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag
     1681 tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt
     1741 tacgtttctc gttcagcttt cttgtacaaa gtggttcgat ctagagggcc cttcgaaggt
     1801 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat
     1861 caccattgag tttaaacccg ctgatcctag agggccgcat catgtaatta gttatgtcac
     1921 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     1981 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     2041 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     2101 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag ctcgaggggg
     2161 ggcccggtac ccaattcgcc ctatagtgag tcgtattacg cgcggatcca gctttggact
     2221 tcttcgccag aggtttggtc aagtctccaa tcaaggttgt cggcttgtct accttgccag
     2281 aaatttacga aaagatggaa aagggtcaaa tcgttggtag atacgttgtt gacacttcta
     2341 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
     2401 gtgtatacaa attttaaagt gactcttagg ttttaaaacg araattctta ttcttgagta
     2461 actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct cttattgacc
     2521 acacctctac cggcatgcca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     2581 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     2641 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2701 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcataatcgg
     2761 atcgtacttg ttacccatca ttgaattttg aacatccgaa cctgggagtt ttccctgaaa
     2821 cagatagtat atttgaacct gtataataat atatagtcta gcgctttacg gaagacaatg
     2881 tatgtatttc ggttcctgga gaaactattg catctattgc ataggtaatc ttgcacgtcg
     2941 catccccggt tcattttctg cgtttccatc ttgcacttca atagcatatc tttgttaacg
     3001 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
     3061 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
     3121 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
     3181 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
     3241 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
     3301 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
     3361 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
     3421 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
     3481 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
     3541 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
     3601 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
     3661 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
     3721 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
     3781 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
     3841 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt
     3901 gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg
     3961 aagttcctat actttctagc tagagaatag gaacttcgga ataggaactt caaagcgttt
     4021 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac
     4081 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
     4141 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct
     4201 agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta
     4261 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct
     4321 atcatttcct ttgatattgg atcgatccga tgataagctg tcaaacatga gaattgggta
     4381 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
     4441 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
     4501 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
     4561 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
     4621 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
     4681 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
     4741 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
     4801 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
     4861 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
     4921 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
     4981 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
     5041 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
     5101 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     5161 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     5221 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg
     5281 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     5341 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     5401 agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa
     5461 atgatgaatt gaaaagctaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     5521 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5581 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5641 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5701 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5761 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5821 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5881 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5941 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6001 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6061 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6121 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6181 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6241 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6301 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6361 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6421 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6481 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6541 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6601 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6661 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6721 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6781 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6841 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6901 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6961 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7021 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7081 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7141 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7201 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7261 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7321 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7381 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7441 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7501 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7561 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7621 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7681 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttcc tgaaacggag
     7741 aaacataaac aggcattgct gggatcaccc atacatcact ctgttttgcc tgaccttttc
     7801 cggtaatttg aaaacaaacc cggtctcgaa gcggagatcc ggcgataatt accgcagaaa
     7861 taaacccata cacgagacgt agaaccagcc gcacatggcc ggagaaactc ctgcgagaat
     7921 ttcgtaaact cgcgcgcatt gcatctgtat ttcctaatgc ggcacttcca ggcctcgatc
     7981 gagaccgttt atccattgct tttttgttgt ctttttccct cgttcacaga aagtctgaag
     8041 aagctatagt agaactatga gctttttttg tttctgtttt cctttttttt ttttttacct
     8101 ctgtggaaat tgttactctc acactcttta gttcgtttgt ttgttttgtt tattccaatt
     8161 atgaccggtg acgaaacgtg gtcgatggtg ggtaccgctt atgctcccct ccattagttt
     8221 cgattatata aaaaggccaa atattgtatt attttcaaat gtcctatcat tatcgtctaa
     8281 catctaattt ctcttaaatt ttttctcttt ctttcctata acaccaatag tgaaaatctt
     8341 tttttcttct atatctacaa aaactttttt tttctatcaa cctcgttgat aaattttttc
     8401 tttaacaatc gttaataatt aattaattgg aaaataacca ttttttctct cttttataca
     8461 cacattcaaa agaaagaaaa aaaatatacc ccagcct
//
