LOCUS       40924_48758        9859 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector SBa_000560, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9859)
  AUTHORS   Temme K, Zhao D, Voigt CA.
  TITLE     Refactoring the nitrogen fixation gene cluster from Klebsiella 
            oxytoca
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 109 (18), 7085-7090 (2012)
  PUBMED    22509035
REFERENCE   2  (bases 1 to 9859)
  AUTHORS   Temme K, Zhao D, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-APR-2012) Biological Engineering, MIT, 500 Technology 
            Square, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 9859)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9859)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2012"; volume: "109"; issue: "18"; pages:
            "7085-7090"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-APR-2012) Biological Engineering, MIT, 500 Technology Square, 
            Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..60
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     terminator      complement(120..149)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    196..217
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     CDS             complement(255..1334)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(1335..1412)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     protein_bind    1963..1979
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    1989..2028
                     /label=SBa_000508
                     /note="SBa_000508"
     regulatory      2029..2062
                     /label=SBa_000507
                     /note="SBa_000507"
                     /regulatory_class="ribosome_binding_site"
     regulatory      2063..2188
                     /label=SBa_000509
                     /note="SBa_000509"
                     /regulatory_class="other"
     CDS             2189..4834
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
     misc_feature    4838..4858
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4894..4925
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     regulatory      4973..5037
                     /label=SBa_000488
                     /note="SBa_000488"
                     /regulatory_class="terminator"
     CDS             5787..6755
                     /label=RepA
                     /note="replication protein of plasmid pSa"
     rep_origin      6795..7230
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             8911..9699
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
ORIGIN
        1 tccggcaaaa aaacgggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga
       61 ttacttcgcg tttgccacct gacgtctaag aaaaggaata ttcagcaatt tgcccgtgcc
      121 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgctacgtaa ttagttagtt
      181 agcccttagt gactcgaatt cgcggccgct tctagagcaa ttcgcgctaa cttacattaa
      241 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
      301 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
      361 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
      421 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttgac
      481 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
      541 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
      601 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
      661 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
      721 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
      781 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
      841 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
      901 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
      961 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     1021 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     1081 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     1141 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     1201 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     1261 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     1321 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     1381 ccgcgaaagg ttttgcacca ttcgatggtg tcaacgtaaa tgcatgccgc ttcgccttcg
     1441 cgcgcgaatt ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     1501 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     1561 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     1621 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     1681 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggcg
     1741 gagcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
     1801 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
     1861 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt aacaccgtgc
     1921 gtgttgacag ctatacctct ggcggttata atgctagcgg aattgtgagc ggataacaat
     1981 ttactagatg cagttttatt ctctcgccag cactgtaata ggcactaata tccaaaccag
     2041 tagctcaatt ggagtcgtct atgtgttgtt tatcaagcct gcggatctcc gcgaaattgt
     2101 gacttttccg ctatttagcg atcttgttca gtgtggcttt ccttcaccgg cagcagatta
     2161 cgttgaacag cgcatcgatc tgggtggcaa cacgattaac atcgctaaga acgacttctc
     2221 tgacatcgaa ctggctgcta tcccgttcaa cactctggct gaccattacg gtgagcgttt
     2281 agctcgcgaa cagttggccc ttgagcatga gtcttacgag atgggtgaag cacgcttccg
     2341 caagatgttt gagcgtcaac ttaaagctgg tgaggttgcg gataacgctg ccgccaagcc
     2401 tctcatcact accctactcc ctaagatgat tgcacgcatc aacgactggt ttgaggaagt
     2461 gaaagctaag cgcggcaagc gcccgacagc cttccagttc ctgcaagaaa tcaagccgga
     2521 agccgtagcg tacatcacca ttaagaccac tctggcttgc ctaaccagtg ctgacaatac
     2581 aaccgttcag gctgtagcaa gcgcaatcgg tcgggccatt gaggacgagg ctcgcttcgg
     2641 tcgtatccgt gaccttgaag ctaagcactt caagaaaaac gttgaggaac aactcaacaa
     2701 gcgcgtaggg cacgtctaca agaaagcatt tatgcaagtt gtcgaggctg acatgctctc
     2761 taagggtcta ctcggtggcg aggcgtggtc ttcgtggcat aaggaagact ctattcatgt
     2821 aggagtacgc tgcatcgaga tgctcattga gtcaaccgga atggttagct tacaccgcca
     2881 aaatgctggc gtagtaggtc aagactctga gactatcgaa ctcgcacctg aatacgctga
     2941 ggctatcgca acccgtgcag gtgcgctggc tggcatctct ccgatgttcc aaccttgcgt
     3001 agttcctcct aagccgtgga ctggcattac tggtggtggc tattgggcta acggtcgtcg
     3061 tcctctggcg ctggtgcgta ctcacagtaa gaaagcactg atgcgctacg aagacgttta
     3121 catgcctgag gtgtacaaag cgattaacat tgcgcaaaac accgcatgga aaatcaacaa
     3181 gaaagtccta gcggtcgcca acgtaatcac caagtggaag cattgtccgg tcgaggacat
     3241 ccctgcgatt gagcgtgaag aactcccgat gaaaccggaa gacatcgaca tgaatcctga
     3301 ggctctcacc gcgtggaaac gtgctgccgc tgctgtgtac cgcaaggaca aggctcgcaa
     3361 gtctcgccgt atcagccttg agttcatgct tgagcaagcc aataagtttg ctaaccataa
     3421 ggccatctgg ttcccttaca acatggactg gcgcggtcgt gtttacgctg tgtcaatgtt
     3481 caacccgcaa ggtaacgata tgaccaaagg actgcttacg ctggcgaaag gtaaaccaat
     3541 cggtaaggaa ggttactact ggctgaaaat ccacggtgca aactgtgcgg gtgtcgacaa
     3601 ggttccgttc cctgagcgca tcaagttcat tgaggaaaac cacgagaaca tcatggcttg
     3661 cgctaagtct ccactggaga acacttggtg ggctgagcaa gattctccgt tctgcttcct
     3721 tgcgttctgc tttgagtacg ctggggtaca gcaccacggc ctgagctata actgctccct
     3781 tccgctggcg tttgacgggt cttgctctgg catccagcac ttctccgcga tgctccgaga
     3841 tgaggtaggt ggtcgcgcgg ttaacttgct tcctagtgaa accgttcagg acatctacgg
     3901 gattgttgct aagaaagtca acgagattct acaagcagac gcaatcaatg ggaccgataa
     3961 cgaagtagtt accgtgaccg atgagaacac tggtgaaatc tctgagaaag tcaagctggg
     4021 cactaaggca ctggctggtc aatggctggc ttacggtgtt actcgcagtg tgactaagcg
     4081 ttcagtcatg acgctggctt acgggtccaa agagttcggc ttccgtcaac aagtgctgga
     4141 agataccatt cagccagcta ttgattccgg caagggtctg atgttcactc agccgaatca
     4201 ggctgctgga tacatggcta agctgatttg ggaatctgtg agcgtgacgg tggtagctgc
     4261 ggttgaagca atgaactggc ttaagtctgc tgctaagctg ctggctgctg aggtcaaaga
     4321 taagaagact ggagagattc ttcgcaagcg ttgcgctgtg cattgggtaa ctcctgatgg
     4381 tttccctgtg tggcaggaat acaagaagcc tattcagacg cgcttgaacc tgatgttcct
     4441 cggtcagttc cgcttacagc ctaccattaa caccaacaaa gatagcgaga ttgatgcaca
     4501 caaacaggag tctggtatcg ctcctaactt tgtacacagc caagacggta gccaccttcg
     4561 taagactgta gtgtgggcac acgagaagta cggaatcgaa tcttttgcac tgattcacga
     4621 ctccttcggt acgattccgg ctgacgctgc gaacctgttc aaagcagtgc gcgaaactat
     4681 ggttgacaca tatgagtctt gtgatgtact ggctgatttc tacgaccagt tcgctgacca
     4741 gttgcacgag tctcaattgg acaaaatgcc agcacttccg gctaaaggta acttgaacct
     4801 ccgtgacatc ttagagtcgg acttcgcgtt cgcgtaatac tagtagcggc cgctgcagga
     4861 gtcactaagg gttagttagt tagattagca gaaagtcaaa agcctccgac cggaggcttt
     4921 tgactaaaac ttcccttggg gttatcattg gggctcactc aaaggcggta atcagataaa
     4981 aaaaatcctt agctttcgct aaggatgatt tctgctagag ctgtcagacc agagctccaa
     5041 accaacgttt tatctatacc gctacagggt atttaattcc tatttaatct gcgctagaat
     5101 gaggcatgtt taaccgaatc tgacgttttc cctgcaaatg ccaaaatact atgcctatct
     5161 ccgggtttcg cgtgacggcc aagacccgga aaaccaaaaa tacggtttgc tcgaatacgc
     5221 gaacgccaaa ggcttcgcgc cgctacagat cgaggaagaa attgccagca gagcaaagga
     5281 ctggcgcaag cgcaagctcg gagcaatcat cgaaaaggcc gagcgtggcg acgtgctact
     5341 gacgccggag attacgcgca ttgccggttc cgccctcgcc gccttggaaa ttctcaaagc
     5401 ggcgagcgag cgcggcctaa tcgtccatgt gaccaaacag aagatcatca tggacggcag
     5461 cctacaaagc gacatcatgg caaccgtgct tggcttggct gcacagatcg agcggcattt
     5521 cattcaggca cgtaccaccg aggcgctaca agtcgccaga gagcgcggca agacgctcgg
     5581 gcgacccaag ggcagcaaat cgagcgcctt gaagctggac agccgtattg atgaagtaca
     5641 ggcatacgtg aaccttggct tgccgcaaag tcgcgcagcc gagttgttag gcgtcagccc
     5701 tcacaccttg cgcctgttca tcaaacgccg gaacatcaaa cccacaaaca ctagaccaac
     5761 catcaccatg ccggggaggg aacaacatgc ctaagaacaa caaagccccc ggccatcgta
     5821 tcaacgagat catcaagacg agcctcgcgc tcgaaatgga ggatgcccgc gaagctggct
     5881 tagtcggcta catggcccgt tgccttgtgc aagcgaccat gccccacacc gaccccaaga
     5941 ccagctactt tgagcgcacc aatggcatcg tcaccttgtc gatcatgggc aagccgagca
     6001 tcggcctgcc ctacggttct atgccgcgca ccttgcttgc ttggatatgc accgaggccg
     6061 tgcgaacgaa agaccccgtg ttgaaccttg gccggtcgca atcggaattt ctacaaaggc
     6121 tcggaatgca caccgatggc cgttacacgg ccacccttcg caatcaggcg caacgcctgt
     6181 tttcatccat gatttcgctt gccggcgagc aaggcaatga cttcggcatt gagaacgtcg
     6241 tcattgccaa gcgcgctttt ctattctgga atcccaagcg gccagaagat cgggcgctat
     6301 gggatagcac cctcaccctc acaggcgatt tcttcgagga agtcacccgc tcaccggttc
     6361 ctatccgaat cgactacctg catgccttgc ggcagtctcc gcttgcgatg gacatttaca
     6421 cgtggctgac ctatcgcgtg ttcctgttgc gggccaaggg ccgccccttc gtgcaaatcc
     6481 cttgggtcgc cctgcaagcg caattcggct catcctatgg cagccgcgca cgcaactcgc
     6541 ccgaactgga cgataaggcc cgagagcggg cagagcgggc agcactcgcc agcttcaaat
     6601 acaacttcaa aaagcgccta cgcgaagtgt tgattgtcta tcccgaggca agcgactgca
     6661 tcgaagatga cggcgaatgc ctgcgcatca aatccacacg cctgcatgtc acccgcgcac
     6721 ccggcaaggg cgctcgcatc ggcccccctc cgacttgacc aggccaacgc tacgcttggc
     6781 ttggtcaagc cttcccatcc aacagcccgc cgtcgagcgg gcttttttat ccccggaagc
     6841 ctgtggatag agggtagtta tccacgtgaa accgctaatg ccccgcaaag ccttgattca
     6901 cggggctttc cggcccgctc caaaaactat ccacgtgaaa tcgctaatca gggtacgtga
     6961 aatcgctaat cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa
     7021 ggcacgtgag aacgctaata gccctttcag atcaacagct tgcaaacacc cctcgctccg
     7081 gcaagtagtt acagcaagta gtatgttcaa ttagcttttc aattatgaat atatatatca
     7141 attattggtc gcccttggct tgtggacaat gcgctacgcg caccggctcc gcccgtggac
     7201 aaccgcaagc ggttgcccac cgtcgagcgc cagcgccttt gcccacaacc cggcggccgc
     7261 aacagatcgt tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct
     7321 ctcgggcttc tggatttccg atcaacgcag gagtcgttcg gaaagtagct gttccagaat
     7381 tataggcgca gagacaccag attccaagat ggctctgtta aattgttgta gtatgtagta
     7441 tcatacaaca tactacagta cagaggcccg caagaatggc aatcactaaa caagacattt
     7501 ggcgagcagc cgacgaactg gacgccgaag gcatccggcc cactttggcc gccgtgcgca
     7561 agaaactcgg aagcggtagc ttcacaacca tttccgatgc aatggctgaa tggaaaaacc
     7621 gcaagaccgc caccctgccc tcatcagacc cattgccggt tgcagtcaac gagcatcttg
     7681 ccgagcttgg caatgcgcta tgggctatcg ccctggcgca cgccaacgcc cggtttgacg
     7741 aagatcggaa acagatcgag gccgacaaag cggccatcag ccagcagctt gccgaagcaa
     7801 tcgaactagc cgacaccttc acccgcgaaa acgaccagct ccgcgaacga gtgaatcagc
     7861 tcgaacctat ggaacgcgag cgcgacaagc tggccgacca gcttgccgaa gtgaaacgcc
     7921 gcagcggcga agaactaaac cgctgcatgg aaaagctcac ccaacgcgat aacgaggcta
     7981 tcgaggcccg caaacaggcc aaggaggcca tcgagcgcgc cgccagtctg caaggtcagg
     8041 tggaagccct caaagagcag gtcgccaatc tcacagccgt cttgaaaaca ggaggcaaac
     8101 aatgaaaagc gcccttgccg cccttcgcgc ggtcgcggcc gctgtcgtcc taatcgtcag
     8161 tgtgcccgct tgggccgact tccggggtga agtcgtccga atccttgacg gtgacactat
     8221 cgacgttttg gtgaaccgtc agaccatccg cgtgagattg gccgatattg acgcaccgga
     8281 aagcggccaa gccttcggct cccgtgctcg ccaacggctc gccgacttga cctttcgcca
     8341 agaggttcaa gtgaccgaaa aagaggttga tcggtatggc cgcactcttg gggtcgttta
     8401 cgcgccgttg caataccccg gcggccaaac acaactcacc aacatcaatg cgatcatggt
     8461 tcaagaaggc atggcctggg cttaccgtta ttacggcaaa ccaaccgacg cgcagatgta
     8521 cgagtatgaa aaagaggccc gccgccaacg gctcggcctt tggtcagacc cgaatgctca
     8581 ggagccttgg aaatggcgtc gcgcctcgaa aaatgccacg aactgacacc gggcacgccc
     8641 cttgttcgac gcgccgcagg cacgtcgaat ttaccgccgg gacgcccctc gtcccgacac
     8701 ttccagatcg ccatagcgca cagcgcctcg agcggtggta acggcgcagt ggcggttttc
     8761 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
     8821 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
     8881 ggcagtcgcc ctaaaacaaa gttaaacatc atgagggaag cggtgatcgc cgaagtatcg
     8941 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     9001 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     9061 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     9121 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     9181 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     9241 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     9301 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     9361 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     9421 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     9481 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
     9541 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
     9601 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
     9661 cactacgtga aaggcgagat caccaaggta gtcggcaaat aatgtctaac aattcgttca
     9721 agccgacgcc gcttcgcggc gcggcttaac tcaagcgtta gatgcactaa gcacataatt
     9781 gctcacagcc aaactatcag gtcaagtctg cttttattat ttttaagcgt gcataataag
     9841 ccctacacaa atggtaccc
//
