LOCUS       62056_23190        8346 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 8346)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             49..75
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     promoter        complement(208..226)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(233..249)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      394..849
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(983..1783)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVLGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVQGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1784..2004)
                     /label=URA3 promoter
     primer_bind     2277..2293
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2853..4195)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        4484..4588
                     /label=AmpR promoter
     CDS             4589..5446
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLARRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLDNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5620..6208
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6496..6517
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6532..6562
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6570..6586
                     /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     6594..6610
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7224..7327)
                     /label=AmpR promoter
     promoter        7835..8345
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose
                     phosphate isomerase"
ORIGIN
        1 gaattcacca tggatcctag ggcccacaag cttacgcgtc gacccgggta tccgtatgat
       61 gtgcctgact acgcatgata tctcgagctc agctagctaa ctgaataagg aacaatgaac
      121 gtttttcctt tctcttgttc ctagtattaa tgactgaccg atacatccct tttttttttt
      181 gtctttgtct agctccagct tttgttccct ttagtgaggg ttaattcaat tcactggccg
      241 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
      301 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
      361 aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg
      421 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
      481 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      541 taaatcgggg gctcccttta gggttccgat ttagtggttt acggcacctc gaccccaaaa
      601 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      661 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      721 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      781 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
      841 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      901 agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat
      961 ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc
     1021 ctgcttttct gtaacgttca ccctgtacct tagcatccct tccctttgca aatagtcctc
     1081 ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt ctatactgtt
     1141 gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc aaccaatcgt
     1201 aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca aaatctttgt
     1261 cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg gagcccttgc
     1321 atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct tctgccgcct
     1381 gcttcaaacc gctaacaata cctggcccca gcacaccgtg tgcattcgta atgtctgccc
     1441 attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta ccaatgtcag
     1501 caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc tttagcggct
     1561 taactgtgcc ctccatcgaa aaatcagtca atatatccac atgtgttttt agtaaacaaa
     1621 ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga acatccaatg
     1681 aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca gcaacaggac
     1741 taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat cttcgtttcc
     1801 tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa
     1861 cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct
     1921 gttcggagat taccgaatca aaaaaatttc aaagaaaccg aaatcaaaaa aaagaataaa
     1981 aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg cactctcagt acaatctgct
     2041 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2101 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2161 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2221 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtgc ggccgctcta
     2281 gaactagtgg atcaattcca cggactatag actatactag tatactccgt ctactgtacg
     2341 atacacttcc gctcaggtcc ttgtccttta acgaggcctt accactcttt tgttactcta
     2401 ttgatccagc tcagcaaagg cagtgtgatc taagattcta tcttcgcgat gtagtaaaac
     2461 tagctagacc gagaaagaga ctagaaatgc aaaaggcact tctacaatgg ctgccatcat
     2521 tattatccga tgtgacgctg cagcttctca atgatattcg aatacgcttt gaggagatac
     2581 agcctaatat ccgacaaact gttttacaga tttacgatcg tacttgttac ccatcattga
     2641 attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat
     2701 aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa
     2761 ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt
     2821 tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag
     2881 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     2941 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     3001 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     3061 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     3121 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     3181 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     3241 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     3301 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     3361 taaaggcatc cccgattata ttctataccg atgtggattc cgcatacttt gtgaacagaa
     3421 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     3481 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     3541 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     3601 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     3661 gggatatagc acagagatat atagcaaaga gatacttttc agcaatgttt gtggaagcgg
     3721 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     3781 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat
     3841 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     3901 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     3961 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     4021 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     4081 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     4141 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatatg
     4201 catagtaccg agaaactagt gcgaagtagt gatcaggtat tgctgttatc tgatgagtat
     4261 acgttgtcct ggccacggca gaagcaccct tatcgctcca atttcccaca acattagtca
     4321 actccgttag gcccttcatt gaaagaaatg aggtcatcaa atgtcttcca atgtgagatt
     4381 ttgggccatt ttttatagca aagattgaat aaggcgcatt tttcttcaaa gctgcggccg
     4441 cactctcact agtacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4501 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cgtgataaat
     4561 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4621 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4681 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4741 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4801 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcgctcg
     4861 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4921 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4981 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgga
     5041 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5101 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5161 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5221 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5281 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5341 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5401 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5461 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5521 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5581 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5641 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5701 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5761 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5821 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5881 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5941 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6001 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6061 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6121 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6181 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6241 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6301 taaccgtatt accgcctttg agtgagctga taccggtcgc cgcagccgaa cgaccgagcg
     6361 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6421 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6481 tgagcgcaac gcaattaatg tgagttacct cactcattag gcaccccagg ctttacactt
     6541 tatgcttccg gctcctatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6601 cagctatgac catgattacg ccaagctcga aatacgactc actatagggc gaattgggta
     6661 ccgggccggc cgtcgagctt gatggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgaaaaa
     6781 aagcggttag ctcttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6841 actcatggtt atggcaggaa ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6901 tttctgtgac tggtgtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6961 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     7021 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7081 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7141 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7201 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7261 ggttattgtc tcatgagcga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7321 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7381 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttggggatct
     7441 acgtatggtc attcttcttc agattccctc atggagaagt gcggcagatg tatatgacag
     7501 agtcgccagt ttccaagaga ctttattcag gcacttccat gataggcaag agagaagacc
     7561 cagagatgtt gttgtcctag ttacacatgg tatttattcc agagtattcc tgatgaaatg
     7621 gtttagatgg acatacgaag agtttgaatc gtttaccaat gttcctaacg ggagcgtaat
     7681 ggtgatggaa ctggacgaat ccatcaatag atacgtcctg aggaccgtgc tacccaaatg
     7741 gactgattgt gagggagacc taactacata gtgtttaaag attacggata tttaacttac
     7801 ttagaataat gccatttttt tgagttataa taatcctacg ttagtgtgag cgggatttaa
     7861 actgtgagga cctcaataca ttcagacact tctgacggta tcaccctact tattcccttc
     7921 gagattatat ctaggaaccc atcaggttgg tggaagatta cccgttctaa gacttttcag
     7981 cttcctctat tgatgttaca ctcggacacc ccttttctgg catccagttt ttaatcttca
     8041 gtggcatgtg agattctccg aaattaatta aagcaatcac acaattctct cggataccac
     8101 ctcggttgaa actgacaggt ggtttgttac gcatgctaat gcaaaggagc ctatatacct
     8161 ttggctcggc tgctgtaaca gggaatataa agggcagcat aatttaggag tttagtgaac
     8221 ttgcaacatt tactattttc ccttcttacg taaatatttt tctttttaat tctaaatcaa
     8281 tctttttcaa ttttttgttt gtattctttt cttgcttaaa tctataacta caaaaaacac
     8341 atacag
//
