LOCUS       62056_19050        3505 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3505)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             169..192
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     CDS             199..849
                     /codon_start=1
                     /label=mKO2
                     /note="monomeric Kusabira-Orange 2 fluorescent protein"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     terminator      920..967
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1064..1519
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1545..1649
                     /label=AmpR promoter
     CDS             1650..2507
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2681..3269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatccgat ggtgagcgtg atcaagcccg agatgaagat gaggtactac
      241 atggacggct ccgtcaatgg gcatgagttc acaatcgagg gtgagggcac aggcagacct
      301 tacgagggac atcaggagat gacactgcgc gtcacaatgg ccgagggcgg gccaatgcct
      361 ttcgccttcg acctggtgtc ccacgtgttc tgttacggcc acagagtttt taccaagtac
      421 ccagaagaga tcccagacta tttcaagcag gcctttcctg agggcctgtc ctgggagagg
      481 tccctggagt tcgaggacgg cggctccgcc tccgtgagcg cccacatcag cctgaggggc
      541 aacaccttct accacaagtc caagttcacc ggcgtgaact tccccgccga cggccccatc
      601 atgcagaacc agagcgtgga ctgggagccc tccaccgaga agatcaccgc cagcgacggc
      661 gtgctgaagg gcgacgtgac catgtacctg aagctggagg gcggcggcaa ccacaagtgc
      721 cagatgaaga ccacctacaa ggccgccaag gagatcctgg agatgcccgg cgaccactac
      781 atcggccaca ggctggtgag gaagaccgag ggcaacatca ccgagcaggt ggaggacgcc
      841 gtggcccact aagcttgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
      901 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
      961 ttttttgctg aaaggaggaa ctatatccgg atctggcgta atagcgaaga ggcccgcacc
     1021 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     1081 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1141 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1201 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1261 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1321 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1381 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1441 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1501 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1561 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1621 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1681 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1741 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1801 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1861 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1921 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1981 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2041 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2101 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2161 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2221 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2281 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2341 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2401 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2461 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2521 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2581 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2641 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2701 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2761 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2821 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2881 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2941 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3001 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3061 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3121 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3181 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3241 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3301 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3361 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3421 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3481 cgcgttggcc gattcattaa tgcag
//
