pAA3.stelN

LOCUS       pAA3.stelN                4639 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses TelN enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    pAA3.stelN
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4639)
  AUTHORS   Jenni S, Harrison SC
  TITLE     Structure of the DASH/Dam1 complex shows its role at the yeast 
            kinetochore-microtubule interface.
  JOURNAL   Science. 2018 May 4;360(6388):552-558. doi: 10.1126/science.aar6436.
  PUBMED    29724956
REFERENCE   2  (bases 1 to 4639)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4639
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    complement(138..171)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     361..380
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        361..379
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             411..433
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             453..476
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             492..515
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             531..554
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             555..572
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             573..593
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             594..2486
                     /codon_start=1
                     /gene="telN"
                     /product="protelomerase of coliphage N15"
                     /label=TelN
                     /note="generates the covalently closed hairpin telomeres"
                     /translation="SKVKIGELINTLVNEVEAIDASDRPQGDKTKRIKAAAARYKNALF
                     NDKRKFRGKGLQKRITANTFNAYMSRARKRFDDKLHHSFDKNINKLSEKYPLYSEELSS
                     WLSMPTANIRQHMSSLQSKLKEIMPLAEELSNVRIGSKGSDAKIARLIKKYPDWSFALS
                     DLNSDDWKERRDYLYKLFQQGSALLEELHQLKVNHEVLYHLQLSPAERTSIQQRWADVL
                     REKKRNVVVIDYPTYMQSIYDILNNPATLFSLNTRSGMAPLAFALAAVSGRRMIEIMFQ
                     GEFAVSGKYTVNFSGQAKKRSEDKSVTRTIYTLCEAKLFVELLTELRSCSAASDFDEVV
                     KGYGKDDTRSENGRINAILAKAFNPWVKSFFGDDRRVYKDSRAIYARIAYEMFFRVDPR
                     WKNVDEDVFFMEILGHDDENTQLHYKQFKLANFSRTWRPEVGDENTRLVALQKLDDEMP
                     GFARGDAGVRLHETVKQLVEQDPSAKITNSTLRAFKFSPTMISRYLEFAADALGQFVGE
                     NGQWQLKIETPAIVLPDEESVETIDEPDDESQDDELDEDEIELDEGGGDEPTEEEGPEE
                     HQPTALKPVFKPAKNNGDGTYKIEFEYDGKHYAWSGPADSPMAAMRSAWETYYS"
     primer_bind     complement(2558..2576)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2572..2619
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2682..2704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2790..2929
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2845..2864
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2944..2961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3115..3703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3195..3214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3734..3773
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(4638..90)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
       61 tatttgaatg tatttagaaa aataaacaaa taggggaatt aaaaaaaagc ccgctcatta
      121 ggcgggctac tagtcagata acttcgtata atgtatgcta tacgaagtta tggtaccgcg
      181 gccgcgtaga ggatctgttg atcagcagtt caacctgttg atagtacttc gttaatacag
      241 atgtaggtgt tggcaccatg cataactata acggtcctaa ggtagcgacc taggttctag
      301 acttcatttc cttctccaga ttaactcaat tccagcattt agaaccctac caagtatcga
      361 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
      421 tttaagaagg agatatacat atgaaatctt cttggagcca tccgcagttt gaaaaaggcg
      481 ccatgaccgg ttggtcccac ccgcagttcg aaaaacgtag cgcgggcagc tggagtcatc
      541 cgcaattcga aaaacaccat caccatcacc atgaaaacct gtacttccaa tccagcaagg
      601 taaaaatcgg tgagttgatc aacacgcttg tgaatgaggt agaggcaatt gatgcctcag
      661 accgcccaca aggcgacaaa acgaagagaa ttaaagccgc agccgcacgg tataagaacg
      721 cgttatttaa tgataaaaga aagttccgtg ggaaaggatt gcagaaaaga ataaccgcga
      781 atacttttaa cgcctatatg agcagggcaa gaaagcggtt tgatgataaa ttacatcata
      841 gctttgataa aaatattaat aaattatcgg aaaagtatcc tctttacagc gaagaattat
      901 cttcatggct ttctatgcct acggctaata ttcgccagca catgtcatcg ttacaatcta
      961 aattgaaaga aataatgccg cttgccgaag agttatcaaa tgtaagaata ggctctaaag
     1021 gcagtgatgc aaaaatagca agactaataa aaaaatatcc agattggagt tttgctctta
     1081 gtgatttaaa cagtgatgat tggaaggagc gccgtgacta tctttataag ttattccaac
     1141 aaggctctgc gttgttagaa gaactacacc agctcaaggt caaccatgag gttctgtacc
     1201 atctgcagct aagccctgcg gagcgtacat ctatacagca acgatgggcc gatgttctgc
     1261 gcgagaagaa gcgtaatgtt gtggttattg actacccaac atacatgcag tctatctatg
     1321 atattttgaa taatcctgcg actttattta gtttaaacac tcgttctgga atggcacctt
     1381 tggcctttgc tctggctgcg gtatcagggc gaagaatgat tgagataatg tttcagggtg
     1441 aatttgccgt ttcaggaaag tatacggtta atttctcagg gcaagctaaa aaacgctctg
     1501 aagataaaag cgtaaccaga acgatttata ctttatgcga agcaaaatta ttcgttgaat
     1561 tattaacaga attgcgttct tgctctgctg catctgattt cgatgaggtt gttaaaggat
     1621 atggaaagga tgatacaagg tctgagaacg gcaggataaa tgctatttta gcaaaagcat
     1681 ttaacccttg ggttaaatca tttttcggcg atgaccgtcg tgtttataaa gatagccgcg
     1741 ctatttacgc tcgcatcgct tatgagatgt tcttccgcgt cgatccacgg tggaaaaacg
     1801 tcgacgagga tgtgttcttc atggagattc tcggacacga cgatgagaac acccagctgc
     1861 actataagca gttcaagctg gccaacttct ccagaacctg gcgacctgaa gttggggatg
     1921 aaaacaccag gctggtggct ctgcagaaac tggacgatga aatgccaggc tttgccagag
     1981 gtgacgctgg cgtccgtctc catgaaaccg ttaagcagct ggtggagcag gacccatcag
     2041 caaaaataac caacagcact ctccgggcct ttaaatttag cccgacgatg attagccggt
     2101 acctggagtt tgccgctgat gcattggggc agttcgttgg cgagaacggg cagtggcagc
     2161 tgaagataga gacacctgca atcgtcctgc ctgatgaaga atccgttgag accatcgacg
     2221 aaccggatga tgagtcccaa gacgacgagc tggatgaaga tgaaattgag ctcgacgagg
     2281 gtggcggcga tgaaccaacc gaagaggaag ggccagaaga acatcagcca actgctctaa
     2341 aacccgtctt caagcctgca aaaaataacg gggacggaac gtacaagata gagtttgaat
     2401 acgatggaaa gcattatgcc tggtccggcc ccgccgatag ccctatggcc gcaatgcgat
     2461 ccgcatggga aacgtactac agctaactcg agagatccgg ctgctaacaa acccgcttaa
     2521 ggaagatggc caacttattt acataaggct gctgccaccg ctgagcaata actagcataa
     2581 ccccttgggg cctctaaacg ggtcttgagg ggttttttgg tttaaaccca tctaattggg
     2641 aattccccct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2701 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2761 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     2821 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccatt
     2881 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     2941 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3001 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3061 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3121 taggttccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3181 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3241 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3301 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3361 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3421 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3481 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3541 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3601 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3661 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3721 ttctaccggt ccgtccaaaa aaaaaggctc caaaaggagc ctttaattgt atcggtttac
     3781 caatgcttaa tcagtgaagc ccctatttca gcaatctgtc tatttcgttc gtccatagtt
     3841 gcctgactac ccgtcgtgta gataactacg atacgggaag gcttaccatc tggtccaagt
     3901 gcggcaatga taccccgtga accacgctct cctgccccag atttatcagc aataaaccag
     3961 ccagcaggaa gtgccgaacg cagtagtggt cctgcaactt tatccgcttc aagccagtct
     4021 attaattgtt gccgtgaagc cagagtaagt agttcgccag ttaatagttt gcgtaatgtt
     4081 gttgccattg ccgcaggcat tgtggtgtct cgctcgtcgt ttggtatggc ttcattcagt
     4141 tctggttccc aacggtcaag gcgagttaca tggtctccca tattgtgcaa aaaagcggtt
     4201 aattccttcg gtccaccgat ggttgtcaga agtaagttcg ccgcagtgtt atcactcatc
     4261 gttatggcgg cactacataa ttctcttacc gtcattccgt ccgtaaggtg cttttctgtc
     4321 actggggagt attcaaccaa gtcattctgc gaatagtgta ttcgtcgtcc gagttgctct
     4381 tgccccgcat ctacacggga taatactgct ccacatagca gaactttgaa agtgctcatc
     4441 attggaaaac gctcttcagg gcgaaaactc tcaaggattt taccgctatt caagtccagt
     4501 tctatgtatc ccactcgtgc ccccaattgg tcctctgcgt cttttacttt cacgagcgtt
     4561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4621 aaatgttgaa tactcatac
//