LOCUS       40924_26949        4945 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKOS405-159, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4945)
  AUTHORS   Reisinger SJ, Patel KG, Santi DV.
  TITLE     Total synthesis of multi-kb DNA sequences from oligonucleotides
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4945)
  AUTHORS   Reisinger SJ, Patel KG, Santi DV.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-NOV-2006) Kosan Biosciences, 3832 Bay Center Place, 
            Hayward, CA 94545, USA
REFERENCE   3  (bases 1 to 4945)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4945)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-NOV-2006) Kosan Biosciences, 3832 Bay Center Place, Hayward, CA 
            94545, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(252..1439)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(1487..1515)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     primer_bind     1708..1724
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1835..1851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1859..1875)
                     /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)."
     promoter        complement(1883..1913)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1928..1949)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2126..2228
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2229..2885
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      complement(3126..3714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3888..4745)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4746..4850)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatactgt agtgatgctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg
      241 gcttccattc aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag
      301 acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg
      361 gcataaatcg ccgtgacgat cagcggtcca gtgatcgaag ttaggctggt aagagccgcg
      421 agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc
      481 aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag
      541 cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata
      601 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
      661 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
      721 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
      781 aagaatacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
      841 actgggttga aggctctcaa gggcatcggt cgacgctctc ccttatgcga ctcctgcatt
      901 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
      961 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     1021 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     1081 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     1141 tagaggatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt
     1201 agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt
     1261 gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg
     1321 ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     1381 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     1441 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     1501 atgataagct gtcaaacatg agaattacaa cttatatcgt atggggcatc atctgctgga
     1561 ctttgcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     1621 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     1681 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctctagcggc
     1741 cgccctcagc tatatcgcta tcgatgagct ctgtagccgt aaagacgtca atgcagcgat
     1801 cactagctga ggttaattaa gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt
     1861 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1921 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1981 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2041 tgcgtattgg gcgctcttcc gctaatcctg gtgtccctgt tgataccggg aagccctggg
     2101 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
     2161 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     2221 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     2281 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     2341 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     2401 cctttattca cattcttgcc cgcctgatga atgctcatcc ggagttccgt atggcaatga
     2461 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     2521 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     2581 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     2641 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     2701 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     2761 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtctgtgatg
     2821 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     2881 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggtgct acgcctgaat
     2941 aactcgagng cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3001 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3061 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3541 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4021 ccgcgagatc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4861 cgaaaagtgc caccttacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4921 aggcgtatca cgaggccctt tcgtc
//
