LOCUS       40924_27628        9609 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Load vector pLD02X, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9609)
  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 9609)
  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 9609)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9609)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Biotechnol."; date: "2014"; volume: "32"; issue: "12"; pages: 
            "1268-1275"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-SEP-2014) Synthetic Biology Center, MIT, 500 Technology Square, 
            NE47, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      34..223
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(466..1054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1228..2085)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2086..2190)
                     /label=AmpR promoter
     rep_origin      complement(2217..3559)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3823..4043
                     /label=URA3 promoter
     CDS             4044..4844
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     rep_origin      complement(4978..5433)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     terminator      complement(5512..5677)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     protein_bind    5918..5938
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     regulatory      5972..6704
                     /regulatory_class="promoter"
     misc_feature    6388..6427
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     misc_feature    6428..6467
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     protein_bind    6716..6740
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     6886..6902
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6907..6931)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(6909..6930)
                     /label=attB2
                     /note="attB2"
     terminator      6954..7141
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    7244..7265
                     /label=attB4
                     /note="attB4"
     protein_bind    7245..7265
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     regulatory      7299..8031
                     /regulatory_class="promoter"
     misc_feature    7715..7754
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     misc_feature    7755..7794
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     protein_bind    8043..8067
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    8043..8064
                     /label=attP1
                     /note="attP1"
     primer_bind     8213..8229
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8234..8258)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      8273..8520
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             8681..9487
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 cctcgagtaa gcttggtacc gcggctagct aagatccgct ctaaccgaaa aggaaggagt
       61 tagacaacct gaagtctagg tccctattta tttttttata gttatgttag tattaagaac
      121 gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat
      181 tatactgaaa accttgcttg agaaggtttt gggacgctcg aagatccagc tgcattaatg
      241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      901 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1201 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1261 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1321 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1381 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1441 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1501 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1561 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1621 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1681 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1741 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1801 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1861 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1921 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1981 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2041 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaacg
     2221 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
     2281 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
     2341 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
     2401 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
     2461 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
     2521 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
     2581 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
     2641 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
     2701 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
     2761 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
     2821 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
     2881 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
     2941 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
     3001 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
     3061 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt
     3121 gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg
     3181 aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga
     3241 aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg
     3301 cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg
     3361 tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta
     3421 cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct
     3481 ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca
     3541 tttcctttga tattggatca tactaagaaa ccattattat catgacatta acctataaaa
     3601 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     3661 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     3721 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     3781 catcagagca gattgtactg agagtgcacc ataccacagc ttttcaattc aattcatcat
     3841 ttttttttta ttcttttttt tgatttcggt ttctttgaaa tttttttgat tcggtaatct
     3901 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     3961 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     4021 cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     4081 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     4141 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     4201 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     4261 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa
     4321 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     4381 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     4441 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     4501 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     4561 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     4621 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat
     4681 tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg
     4741 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     4801 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     4861 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     4921 taccctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     4981 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5041 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5101 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5161 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5221 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5281 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5341 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5401 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     5461 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgaatt ggagcgacct
     5521 catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag aataagaatt
     5581 ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt tttataactt
     5641 atttaataat aaaaatcata aatcataaga aattcgctta tttagaagtg tcaacaacgt
     5701 atctaccaac gatttgaccc ttttccatct tttcgtaaat ttctggcaag gtagacaagc
     5761 cgacaacctt gattggagac ttgaccaaac ctctggcgaa gaattgttaa ttaagagctc
     5821 tggaggttta ccgagctctt attggttttc aaacttcatt gactgtgccg tttaccgagc
     5881 tcttattggt tttcaaactt cattgactgt gccgagccaa ctttgtatag aaaagttggc
     5941 tccgaatttc tcgagggcca gtgccaagct ttctttttta ctttaatatt atgtatactg
     6001 aaaaatgcac gtgatgataa gactttggaa atttgtgtaa aacccccatt tttttttgct
     6061 gctgctattg ccaaagncaa caagtcttca ggagacatca acactaagtt tctaccccgt
     6121 cttcccctag aattctttct gtacgctcag ggtgggcctt taaaggatag caccctaccg
     6181 aagtcgactt ctaagtaaac accattacta ggagatgact aaatctggaa aacacatggt
     6241 ggtctgaatg cgtctagtct ctgccataaa cataacatgt ttgttttaat gcattctcgt
     6301 gtttaatcga cattaatgtg ccgggagaaa gacatcccat ccctgaaagg tttttccagg
     6361 gaatagtcag gacgcattgg ctttcactcg aatttggccg gcccacaccg gtagatttgg
     6421 ctgggcctcg aatttggccg gcccacaccg gtagatttgg ctgggcctcg agaagaagaa
     6481 aggaagacat gtattacatt atccaacaaa aaatggttct tgacgtctac aaatcaagaa
     6541 tcttaaagac attgaacgaa gtagctgaat aaaaattatg aaaactataa aaactataaa
     6601 aactgtactt aagtcctcaa taaaacataa acttcttact gtataaggtt ttcgataatt
     6661 tcttacttga ttctaggaga gcaacggtaa taaaagcaac gacggggaat tcgacccaag
     6721 tttgtacaaa aaagcaggct gagcgccatt cgccattcag gctgcgcaac tgttgggaag
     6781 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6841 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6901 gtgagtaccc agctttcttg tacaaagtgg tctaccgcgc cacttctaaa taagcgaatt
     6961 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     7021 tttaaagtga ctcttaggtt ttaaaacgaa aattcttgtt cttgagtaac tctttcctgt
     7081 aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg
     7141 gcagatcctg aggtgcgttt ttatgcttgt agtattgtat aatgttttta agatccgtgc
     7201 gtttttatgc ttgtagtatt gtataatgtt tttaagatcc gagccaactt tgtatagaaa
     7261 agttggctcc gaatttctcg agggccagtg ccaagctttc ttttttactt taatattatg
     7321 tatactgaaa aatgcacgtg atgataagac tttggaaatt tgtgtaaaac ccccattttt
     7381 ttttgctgct gctattgcca aagncaacaa gtcttcagga gacatcaaca ctaagtttct
     7441 accccgtctt cccctagaat tctttctgta cgctcagggt gggcctttaa aggatagcac
     7501 cctaccgaag tcgacttcta agtaaacacc attactagga gatgactaaa tctggaaaac
     7561 acatggtggt ctgaatgcgt ctagtctctg ccataaacat aacatgtttg ttttaatgca
     7621 ttctcgtgtt taatcgacat taatgtgccg ggagaaagac atcccatccc tgaaaggttt
     7681 ttccagggaa tagtcaggac gcattggctt tcactcgaat ttggccggcc cacaccggta
     7741 gatttggctg ggcctcgaat ttggccggcc cacaccggta gatttggctg ggcctcgaga
     7801 agaagaaagg aagacatgta ttacattatc caacaaaaaa tggttcttga cgtctacaaa
     7861 tcaagaatct taaagacatt gaacgaagta gctgaataaa aattatgaaa actataaaaa
     7921 ctataaaaac tgtacttaag tcctcaataa aacataaact tcttactgta taaggttttc
     7981 gataatttct tacttgattc taggagagca acggtaataa aagcaacgac ggggaattcg
     8041 acccaagttt gtacaaaaaa gcaggctgag cgccattcgc cattcaggct gcgcaactgt
     8101 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     8161 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     8221 acggccagtg agtacccagc tttcttgtac aaagtggtct acgagggccg catcatgtaa
     8281 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
     8341 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     8401 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     8461 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     8521 ggccctgagg tctaataccc aatctctcgt cttatccaga tgttttatac gcctctggcc
     8581 cgtgtctcaa aatctctgat gttacattgc acaagataaa ataatatcat catgaacaat
     8641 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     8701 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     8761 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     8821 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     8881 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     8941 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     9001 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     9061 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     9121 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     9181 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     9241 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     9301 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     9361 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     9421 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     9481 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     9541 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     9601 agcgtcaga
//
