LOCUS       Exported               12242 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Load driver vector pLDUNB, complete sequence.
ACCESSION   KM457489
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12242)
  AUTHORS   Mishra D, Rivera PM, Lin A, Del Vecchio D, Weiss R.
  TITLE     A load driver device for engineering modularity in biological 
            networks
  JOURNAL   Nat. Biotechnol. 32 (12), 1268-1275 (2014)
  PUBMED    25419739
REFERENCE   2  (bases 1 to 12242)
  AUTHORS   Mishra D, Rivera-Ortiz PM, Lin A, Del Vecchio D, Weiss R.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-SEP-2014) Synthetic Biology Center, MIT, 500 
            Technology Square, NE47, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 12242)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12242
                     /organism="Load driver vector pLDUNB"
                     /mol_type="other DNA"
                     /db_xref="taxon:1565118"
     misc_feature    50..542
                     /label=HO-R
                     /note="HO-R"
     promoter        569..599
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             653..1312
                     /codon_start=1
                     /gene="cmR"
                     /product="Cm(R)"
                     /label=cmR
                     /protein_id="AIU94983.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            653..1312
                     /gene="cmR"
                     /label=cmR
     rep_origin      complement(1317..2257)
                     /direction=LEFT
     rep_origin      1487..2075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2383..2866
                     /label=HO-L
                     /note="HO-L"
     misc_feature    2965..2986
                     /label=attB4
                     /note="attB4"
     protein_bind    2966..2986
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      3036..3443
                     /regulatory_class="promoter"
     promoter        3041..3439
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3443..3459
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3485..3509
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3485..3506
                     /label=attP1
                     /note="attP1"
     CDS             3512..4519
                     /codon_start=1
                     /gene="rtTA"
                     /product="rtTA"
                     /label=rtTA
                     /protein_id="AIU94979.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLKGESWQDFLRNNAKSFRCALLSHRNGAKVHSDTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     gene            3512..4519
                     /gene="rtTA"
                     /label=rtTA
     protein_bind    complement(4521..4545)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(4523..4544)
                     /label=attB2
                     /note="attB2"
     regulatory      4550..4761
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     terminator      4568..4755
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    4858..4879
                     /label=attB4
                     /note="attB4"
     protein_bind    4859..4879
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    4910..5228
                     /label=7xTetO
                     /note="7xTetO"
     protein_bind    4933..5203
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     regulatory      5229..5385
                     /regulatory_class="promoter"
     protein_bind    5395..5419
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5396..5416
                     /label=attP1
                     /note="attP1"
     CDS             5428..7296
                     /codon_start=1
                     /gene="Skn7m"
                     /product="SKN7m"
                     /label=Skn7m
                     /note="SKN7 with D427E mutation"
                     /protein_id="AIU94978.1"
                     /translation="MSFSTINSNVNKTTGDSNNNTTENSSTADLLGMDLLQSGPRLMNT
                     MQPNNSSDMLHINNKTNNVQQPAGNTNISSANAGAKAPANEFVRKLFRILENNEYPDIV
                     TWTENGKSFVVLDTGKFTTHILPNHFKHSNFASFVRQLNKYDFHKVKRSPEERQRCKYG
                     EQSWEFQHPEFRVHYGKGLDNIKRKIPAQRKVLLDESQKALLHFNSEGTNPNNPSGSLL
                     NESTTELLLSNTVSKDAFGNLRRRVDKLQKELDMSKMESYATKVELQKLNSKYNTVIES
                     LITFKTINENLLNNFNTLCSTLANNGIEVPIFGDNGNRNPTGNTNPATTTAIQSNNNTN
                     NASPATSTVSLQLPNLPDQNSLTPNAQNNTVTLRKGFHVLLVEDDAVSIQLCSKFLRKY
                     GCTVQVVSDGLSAISTLEKYRYDLVLMEIVMPNLDGATATSIVRSFDNETPIIAMTGNI
                     MNQDLITYLQHGMNDILAKPFTRDDLHSILIRYLKDRIPLCEQQLPPRNSSPQTHSNTN
                     TANSNPNTINEQSLAMLPQDNPSTTTPVTPGASISSAQHVQQGQQEQQHQIFHAQQQQQ
                     HHNAIANARSDVAIPNLEHEINTVPHSSMGSTPQLPQSTLQENQLS"
     gene            5428..7296
                     /gene="Skn7m"
                     /label=Skn7m
     protein_bind    complement(7301..7325)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(7303..7324)
                     /label=attB2
                     /note="attB2"
     regulatory      7330..7591
                     /regulatory_class="terminator"
                     /note="CYC1 terminator"
     terminator      7340..7587
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    7688..7709
                     /label=attB4
                     /note="attB4"
     protein_bind    7689..7709
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      7743..8475
                     /regulatory_class="promoter"
     misc_feature    8159..8198
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     misc_feature    8199..8238
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     protein_bind    8487..8511
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    8487..8508
                     /label=attP1
                     /note="attP1"
     CDS             8519..9235
                     /codon_start=1
                     /gene="yEGFP"
                     /product="EGFP"
                     /label=yEGFP
                     /protein_id="AIU94982.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            8519..9235
                     /gene="yEGFP"
                     /label=yEGFP
     protein_bind    complement(9240..9264)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(9242..9263)
                     /label=attB2
                     /note="attB2"
     terminator      9274..9471
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     regulatory      9277..9501
                     /regulatory_class="terminator"
                     /note="TEF terminator"
     misc_feature    9640..9661
                     /label=attB4
                     /note="attB4"
     protein_bind    9641..9661
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      9711..10118
                     /regulatory_class="promoter"
     promoter        9716..10114
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     10118..10134
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    10160..10184
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    10160..10181
                     /label=attP1
                     /note="attP1"
     CDS             10193..11002
                     /codon_start=1
                     /gene="kanMX"
                     /product="KanMX"
                     /label=kanMX
                     /protein_id="AIU94981.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            10193..11002
                     /gene="kanMX"
                     /label=kanMX
     protein_bind    complement(11004..11028)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(11006..11027)
                     /label=attB2
                     /note="attB2"
     regulatory      11033..11157
                     /regulatory_class="terminator"
                     /note="PHO5 Terminator"
     CDS             11314..12123
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /protein_id="AIU94980.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            11314..12123
                     /gene="kanR"
                     /label=kanR