LOCUS       Exported               10135 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Tri-shuttle cloning vector pmycYACTn, complete sequence.
ACCESSION   GU593054
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10135)
  AUTHORS   Benders GA, Noskov VN, Denisova EA, Lartigue C, Gibson DG, 
            Assad-Garcia N, Chuang RY, Carrera W, Moodie M, Algire MA, Phan Q, 
            Alperovich N, Vashee S, Merryman C, Venter JC, Smith HO, Glass JI, 
            Hutchison CA III.
  TITLE     Cloning whole bacterial genomes in yeast
  JOURNAL   Nucleic Acids Res. 38 (8), 2558-2569 (2010)
  PUBMED    20211840
REFERENCE   2  (bases 1 to 10135)
  AUTHORS   Benders GA, Gibson DG, Lartigue C, Glass JI.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2010) Synthetic Biology, J. Craig Venter 
            Institute, 10355 Science Center Dr., San Diego, CA 92121, USA
REFERENCE   3  (bases 1 to 10135)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10135
                     /organism="Tri-shuttle cloning vector pmycYACTn"
                     /mol_type="other DNA"
                     /db_xref="taxon:747761"
     CDS             84..1256
                     /codon_start=1
                     /product="IS256 transposase"
                     /label=IS256 transposase
                     /protein_id="ADE19325.1"
                     /translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
                     IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
                     ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
                     LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
                     GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
                     FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
                     ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
     misc_feature    1257..3102
                     /label=pBS+ vector sequence
                     /note="pBS+ vector sequence"
     CDS             1327..2187
                     /codon_start=1
                     /gene="bla"
                     /product="bla"
                     /label=bla
                     /protein_id="ADE19328.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1327..2187
                     /gene="bla"
                     /label=bla
     rep_origin      2358..2946
                     /label=pUC19 origin
                     /note="pUC19 origin"
     mobile_element  3103..3128
                     /mobile_element_type="insertion sequence:IS256"
                     /note="outer inverted repeat"
     CDS             complement(3135..6218)
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /protein_id="ADE19329.1"
                     /translation="MDTSKELLKRDPVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE
                     ARTDRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIY
                     TNVTYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWV
                     GYGQDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKP
                     TTQISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGE
                     IIDERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVR
                     IENGLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNH
                     PLWYTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVI
                     IWSLGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFP
                     AVPKWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVD
                     QSLIKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRL
                     SGQTIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPES
                     AGQLWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCI
                     ELGNKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVE
                     RWKAAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAIT
                     VDVEVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMY
                     TPYVFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLN
                     IDGFHMGIGGDDSWSPSVSAELQLSAGRYHYQLVWCQK"
     gene            complement(3135..6218)
                     /gene="lacZ"
                     /label=lacZ
     CDS             complement(3135..6182)
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAELQLSAGRYHYQLVWCQK"
     regulatory      complement(6219..6463)
                     /regulatory_class="promoter"
                     /note="spiralin promoter"
     gene            complement(6464..8428)
                     /gene="tetM"
                     /label=tetM
     CDS             complement(6494..8428)
                     /codon_start=1
                     /gene="tetM"
                     /product="tetM"
                     /label=tetM
                     /protein_id="ADE19326.1"
                     /translation="MEENHMKIINIGVLAHVDAGKTTLTESLLYNSGAITELGSVDKGT
                     TRTDNTLLERQRGITIQTGITSFQWENTKVNIIDTPGHMDFLAEVYRSLSVLDGAILLI
                     SAKDGVQAQTRILFHALRKMGIPTIFFINKIDQNGIDLSTVYQDIKEKLSAEIVIKQKV
                     ELYPNVCVTNFTESEQWDTVIEGNDDLLEKYMSGKSLEALELEQEESIRFQNCSLFPLY
                     HGSAKSNIGIDNLIEVITNKFYSSTHRGPSELCGNVFKIEYTKKRQRLAYIRLYSGVLH
                     LRDSVRVSEKEKIKVTEMYTSINGELCKIDRAYSGEIVILQNEFLKLNSVLGDTKLLPQ
                     RKKIENPHPLLQTTVEPSKPEQREMLLDALLEISDSDPLLRYYVDSTTHEIILSFLGKV
                     QMEVISALLQEKYHVEIEITEPTVIYMERPLKNAEYTIHIEVPPNPFWASIGLSVSPLP
                     LGSGMQYESSVSLGYLNQSFQNAVMEGIRYGCEQGLYGWNVTDCKICFKYGLYYSPVST
                     PADFRMLAPIVLEQVLKKAGTELLEPYLSFKIYAPQEYLSRAYNDAPKYCANIVDTQLK
                     NNEVILSGEIPARCIQEYRSDLTFFTNGRSVCLTELKGYHVTTGEPVCQPRRPNSRIDK
                     VRYMFNKIT"
     regulatory      complement(8435..8757)
                     /regulatory_class="promoter"
                     /note="spiralin promoter"
     misc_feature    8758..9261
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_feature    8758..8874
                     /label=CEN6
                     /note="CEN6"
     rep_origin      8888..9262
                     /label=ARSH4
                     /note="ARSH4"
     promoter        9262..9449
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     regulatory      9263..9423
                     /regulatory_class="promoter"
                     /note="HIS3 promoter"
     CDS             9450..10109
                     /codon_start=1
                     /gene="HIS3"
                     /product="HIS3"
                     /label=HIS3
                     /protein_id="ADE19327.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     gene            9450..10109
                     /gene="HIS3"
                     /label=HIS3
     mobile_element  complement(10110..10135)
                     /mobile_element_type="insertion sequence:IS256"
                     /note="inner inverted repeat"