LOCUS       40924_17434        4686 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEnEOTG-stop, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4686)
  AUTHORS   Koiwa H.
  TITLE     Arabidopsis CPL4 is an essential CTD phosphatase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4686)
  AUTHORS   Koiwa H.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-AUG-2013) Department of Horticultural Science, Texas A
REFERENCE   3  (bases 1 to 4686)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4686)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-AUG-2013) Department of Horticultural Science, Texas A"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..773
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     misc_feature    777..852
                     /label=Tobacco mosaic virus omega
                     /note="Tobacco mosaic virus omega"
     CDS             889..1062
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1066..1236
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             1273..1293
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             1300..1377
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1378..1392
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     primer_bind     complement(1398..1414)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1417..2130
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2179..2431
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    complement(2452..2551)
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      complement(2601..2628)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(2720..2806)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(2971..3559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3652..4458)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     protein_bind    4581..4680
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tagcttgcat gcctgcaggt cgaagcttga acatggtgga gcacgacact ctcgtctact
       61 ccaagaatat caaagataca gtctcagaag accaaagggc tattgagact tttcaacaaa
      121 gggtaatatc gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcaaaa
      181 ggacagtaga aaaggaaggt ggcacctaca aatgccatca ttgcgataaa ggaaaggcta
      241 tcgttcaaga tgcctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca
      301 tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgataaca
      361 tggtggagca cgacactctc gtctactcca agaatatcaa agatacagtc tcagaagacc
      421 aaagggctat tgagactttt caacaaaggg taatatcggg aaacctcctc ggattccatt
      481 gcccagctat ctgtcacttc atcaaaagga cagtagaaaa ggaaggtggc acctacaaat
      541 gccatcattg cgataaagga aaggctatcg ttcaagatgc ctctgccgac agtggtccca
      601 aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt
      661 caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact
      721 atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag aggtacgtat
      781 ttttacaaca attaccaaca acaacaacaa acaacaacaa cattacattt tacattctac
      841 aactacatct agtcgaccat ggcaggcctt gcgcaacacg atgaagccgt ggacaacaaa
      901 ttcaacaaag aacaacaaaa cgcgttctat gagatcttac atttacctaa cttaaacgaa
      961 gaacaacgaa acgccttcat ccaaagttta aaagatgacc caagccaaag cgctaacctt
     1021 ttagcagaag ctaaaaagct aaatgatgct caggcgccga aagtagacaa caaattcaac
     1081 aaagaacaac aaaacgcgtt ctatgagatc ttacatttac ctaacttaaa cgaagaacaa
     1141 cgaaacgcct tcatccaaag tttaaaagat gacccaagcc aaagcgctaa ccttttagca
     1201 gaagctaaaa agctaaatga tgctcaggcg ccgaaagtag acgcgaattg tgatatacct
     1261 acaactgctt ctgaaaattt atattttcaa ggtgaactaa agagaagatg gaaaaagaat
     1321 ttcatagccg tctcagcagc caaccgcttt aagaaaatct catcctccgg ggcacttgat
     1381 gatgacgata agcttatcga taccgtcgac ctcgaggtga gcaagggcga ggagctgttc
     1441 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1501 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1561 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcac ctacggcgtg
     1621 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1681 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1741 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1801 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1861 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1921 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1981 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     2041 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2101 atcactcacg gcatggacga gctgtacaaa acccgggaaa tagagcggcc gccaccgcgg
     2161 tggagctcga atttccccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     2221 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
     2281 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     2341 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     2401 tcgcgcgcgg tgtcatctat gttactagat cgggaattag gttccggcgc cagcctgctt
     2461 ttttgtacaa agttggcatt ataaaaaagc attgcttatc aatttgttgc aacgaacagg
     2521 tcactatcag tcaaaataaa atcattattt ggggcccgag cttaagtaac taactaacag
     2581 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
     2641 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
     2701 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     2761 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
     2821 cctactctcg cttagtagtt agacgtcccc gagatccatg ctagcggtaa tacggttatc
     2881 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2941 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3001 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3421 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3481 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3541 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3601 gaacggggcc caatctgaat aatgttacaa ccaattaacc aattctgatt agaaaaactc
     3661 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     3721 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     3781 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     3841 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     3901 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     3961 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     4021 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     4081 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     4141 ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     4201 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     4261 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     4321 atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     4381 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc
     4441 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     4501 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacggg
     4561 ccagagctgc agctggatgg caaataatga ttttattttg actgatagtg acctgttcgt
     4621 tgcaacaaat tgataagcaa tgctttctta taatgccaac tttgtacaag aaagctgggt
     4681 ctagat
//
