LOCUS       40924_48753        5208 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector SBa_000559, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5208)
  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 5208)
  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 5208)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5208)
  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..5208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    43..64
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     promoter        64..348
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     protein_bind    377..395
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             456..1166
                     /codon_start=1
                     /label=lambda repressor
                     /note="phage lambda repressor"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             1167..1199
                     /codon_start=1
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /translation="AANDENYALVA"
     terminator      1247..1318
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1334..1361
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1435..2310
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             2340..2960
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline repressor TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     misc_feature    2967..2987
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2988..3047
                     /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."
     rep_origin      complement(3294..3881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3969..4063)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4087..4944)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4945..5016)
                     /label=AmpR promoter
     terminator      complement(join(5205..5208,1..40))
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct
       61 agagagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc
      121 tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga
      181 ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca
      241 ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag
      301 cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttt
      361 gggctagcga ctagagtccc tatcagtgat agagattgac atccctatca gtgatagata
      421 taatgagcac tactagagtc acacaggact actagatgag cacaaaaaag aaaccattaa
      481 cacaagagca gcttgaggac gcacgtcgcc ttaaagcaat ttatgaaaaa aagaaaaatg
      541 aacttggctt atcccaggaa tctgtcgcag acaagatggg gatggggcag tcaggcgttg
      601 gtgctttatt taatggcatc aatgcattaa atgcttataa cgccgcattg cttgcaaaaa
      661 ttctcaaagt tagcgttgaa gaatttagcc cttcaatcgc cagagaaatc tacgagatgt
      721 atgaagcggt tagtatgcag ccgtcactta gaagtgagta tgagtaccct gttttttctc
      781 atgttcaggc agggatgttc tcacctgagc ttagaacctt taccaaaggt gatgcggaga
      841 gatgggtaag cacaaccaaa aaagccagtg attctgcatt ctggcttgag gttgaaggta
      901 attccatgac cgcaccaaca ggctccaagc caagctttcc tgacggaatg ttaattctcg
      961 ttgaccctga gcaggctgtt gagccaggtg atttctgcat agccagactt gggggtgatg
     1021 agtttacctt caagaaactg atcagggata gcggtcaggt gtttttacaa ccactaaacc
     1081 cacagtaccc aatgatccca tgcaatgaga gttgttccgt tgtggggaaa gttatcgcta
     1141 gtcagtggcc tgaagagacg tttggcgctg caaacgacga aaactacgct ttagtagctt
     1201 aataacgctg atagtgctag tgtagatcgc tactagagcc aggcatcaaa taaaacgaaa
     1261 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
     1321 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatacta gagttgacag
     1381 ctagctcagt cctagggatt gtgctagcta ctagagaaag aggagaaata ctagatggct
     1441 gaagcgcaaa atgatcccct gctgccggga tactcgttta atgcccatct ggtggcgggt
     1501 ttaacgccga ttgaggccaa cggttatctc gattttttta tcgaccgacc gctgggaatg
     1561 aaaggttata ttctcaatct caccattcgc ggtcaggggg tggtgaaaaa tcagggacga
     1621 gaatttgttt gccgaccggg tgatattttg ctgttcccgc caggagagat tcatcactac
     1681 ggtcgtcatc cggaggctcg cgaatggtat caccagtggg tttactttcg tccgcgcgcc
     1741 tactggcatg aatggcttaa ctggccgtca atatttgcca atacggggtt ctttcgcccg
     1801 gatgaagcgc accagccgca tttcagcgac ctgtttgggc aaatcattaa cgccgggcaa
     1861 ggggaagggc gctattcgga gctgctggcg ataaatctgc ttgagcaatt gttactgcgg
     1921 cgcatggaag cgattaacga gtcgctccat ccaccgatgg ataatcgggt acgcgaggct
     1981 tgtcagtaca tcagcgatca cctggcagac agcaattttg atatcgccag cgtcgcacag
     2041 catgtttgct tgtcgccgtc gcgtctgtca catcttttcc gccagcagtt agggattagc
     2101 gtcttaagct ggcgcgagga ccaacgtatc agccaggcga agctgctttt gagcaccacc
     2161 cggatgccta tcgccaccgt cggtcgcaat gttggttttg acgatcaact ctatttctcg
     2221 cgggtattta aaaaatgcac cggggccagc ccgagcgagt tccgtgccgg ttgtgaagaa
     2281 aaagtgaatg atgtagccgt caagttgtca taatactaga tcacacagga aaggcctcga
     2341 tgtccagatt agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg
     2401 gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat
     2461 tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag
     2521 ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta
     2581 ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt
     2641 taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat
     2701 gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg gggcatttta
     2761 ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac
     2821 ctactactga tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag
     2881 gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac
     2941 ttaaatgtga aagtgggtcc taataatact agtagcggcc gctgcagtcc ggcaaaaaaa
     3001 cgggcaaggt gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta
     3061 tgcaggcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3121 cagctcactc aaaggcggta atcctagggc gttcggctgc ggcgagcggt atcagctcac
     3181 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3481 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3721 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttg
     3841 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3901 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgactt
     3961 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     4021 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc
     4081 aaattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4141 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4201 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4261 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4321 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4381 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4441 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4501 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4561 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4621 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4681 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     4741 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4801 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4861 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4921 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4981 gggttattgt ctcatgagcg gatacatatt tgaatgtatt taggatcaac gtctcatttt
     5041 cgccagatat cgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5101 gtatcacgag gccctttcgt cttcacctcg agtgccacct gacgtctaag aaaccattat
     5161 tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaattt
//
