LOCUS       62056_16635        5972 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 5972)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..937
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             949..1215
                     /codon_start=1
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     CDS             1273..1302
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             1318..1335
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1415..1661
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             1804..2616
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      2692..2939
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2985..3001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3475..3579
                     /label=AmpR promoter
     CDS             3580..4437
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4611..5199
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5487..5508
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5523..5553
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5561..5577
                     /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)."
     primer_bind     5585..5601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaagga tccaacatat gagatttcct
      961 tcaattttta ctgcagtttt attcgcagca tcctccgcat tagctgctcc agtcaacact
     1021 acaacagaag atgaaacggc acaaattccg gctgaagctg tcatcggtta ctcagattta
     1081 gaaggggatt tcgatgttgc tgttttgcca ttttccaaca gcacaaataa cgggttattg
     1141 tttataaata ctactattgc cagcattgct gctaaagaag aaggggtatc tctcgagaaa
     1201 agagaggctg aagcttacgt agaattctcg cgggccccct ggtaccgaca gccgcggtca
     1261 gcggccgctc tagaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat
     1321 catcatcatc atcattgagt ttgtagcctt agacatgact gttcctcagt tcaagttggg
     1381 cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgtcatttg
     1441 cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga
     1501 ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag
     1561 ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg
     1621 tatttcccac tcctcttcag aatacagaag attaagtgag accttcgttt gtgcggatcc
     1681 gctagcttag tgtactaccc aatcttgcct cttcgggtgt cttttataag gacagattca
     1741 caagctcttg ttgcccaata cacacataca cacagagata atagcagtct taattaaatg
     1801 aaaatgagcc atattcaacg ggaaacgtct tgctcaaggc cgcgattaaa ttccaacatg
     1861 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1921 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1981 agcgttgcta atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2041 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2101 gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2161 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2221 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2281 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2341 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2401 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2461 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2521 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2581 aaattgcagt ttcatttgat gctcgatgag tttttctaaa gatctcacgt ccgacgggcg
     2641 gcccacgggt cccaggcctc ggagatccgt cccccttttc ctttgtcgat atcatgtaat
     2701 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc aaaaaggaag
     2761 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     2821 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     2881 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca
     2941 agctagtact tttaatcgtc cacttgccgt catgcttctt tggcactggc cgtcgtttta
     3001 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3061 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3121 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3181 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3241 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3301 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3361 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3421 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3481 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3541 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3601 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3661 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3721 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3781 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3841 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3901 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3961 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4021 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4081 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4141 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4201 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4261 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4321 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4381 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4441 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4501 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4561 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4621 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4681 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4741 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4801 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4861 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4921 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4981 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5041 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5101 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5161 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5221 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5281 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5341 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     5401 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5461 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     5521 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     5581 cacacaggaa acagctatga ccatgattac gaactcgtcc tcggttcccg gcgatcctct
     5641 ggagattgct taccaaaaat ggcccactga gggggtctag tatttgaaga agcctcacta
     5701 aagcaggtct tcgccacaca tttaaagttt gagaatgggc ggaggtcaac cccccgtccc
     5761 cctaatcatt gctttacctc atggaactaa acaaccaccc ccccgctatc ctgagggaaa
     5821 cttcggcagg aaccagctac tagatggttc gattagtctt tcgcccctat acccaaattc
     5881 gacgatcgat ttgcacgtca gaaccgctgc gggcctccac cagagtttcc tcaggcttcg
     5941 ccctattcag gcatagttca ccatctacta gt
//
