LOCUS       40924_27618        5922 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Load vector pLD00X, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5922)
  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 5922)
  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 5922)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5922)
  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..5922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        196..416
                     /label=URA3 promoter
     CDS             417..1217
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     terminator      complement(1885..2050)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     terminator      2329..2518
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(2761..3349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3523..4380)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4381..4485)
                     /label=AmpR promoter
     rep_origin      complement(4512..5854)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1861 ttcgctatta cgccagctga attggagcga cctcatgcta tacctgagaa agcaacctga
     1921 cctacaggaa agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt
     1981 aaaatttgta tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata
     2041 agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc aacgatttga cccttttcca
     2101 tcttttcgta aatttctggc aaggtagaca agccgacaac cttgattgga gacttgacca
     2161 aacctctggc gaagaattgt taattaagag ctctggaggt ttaccgagct cttattggtt
     2221 ttcaaacttc attgactgtg ccggccggcc ggtatcacta gtattacaga ggtaagttat
     2281 aacagtcgcc taacccctcg agtaagcttg gtaccgcggc tagctaagat ccgctctaac
     2341 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     2401 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     2461 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaagat
     2521 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2581 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2641 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2701 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2761 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2881 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2941 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3001 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3061 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3181 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3421 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3481 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3601 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3661 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3721 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3781 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3841 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3901 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3961 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4021 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4081 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4141 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4201 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4261 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4321 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4381 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4441 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4501 agtgccacct gaacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     4561 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     4621 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     4681 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     4741 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     4801 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     4861 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     4921 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     4981 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     5041 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     5101 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     5161 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     5221 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     5281 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     5341 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
     5401 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt
     5461 ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact
     5521 tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc
     5581 tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga
     5641 acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg
     5701 aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc
     5761 cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat
     5821 ccttcaatgc tatcatttcc tttgatattg gatcatacta agaaaccatt attatcatga
     5881 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
