LOCUS       62056_21115        8356 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 8356)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(192..780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(954..1811)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1812..1916)
                     /label=AmpR promoter
     primer_bind     2788..2804
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2811..2829
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(3438..4794)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        4871..5275
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        5293..5936
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     CDS             6230..6940
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      6963..7152
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8144..8160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8168..8184)
                     /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)."
     protein_bind    complement(8239..8260)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
       61 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      121 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      181 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      241 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      301 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      361 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      421 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgtgcc
      481 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      541 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      601 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      661 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      721 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      781 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      841 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      901 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      961 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1021 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1081 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1141 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1201 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1261 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1321 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1381 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1441 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1501 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1561 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1621 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1681 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1741 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1801 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1861 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1921 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1981 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     2041 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     2101 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     2161 gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg
     2221 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc
     2281 ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt
     2341 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     2401 tatcagggcg atggcccacc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2461 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc
     2521 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat
     2581 ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2641 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     2701 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     2761 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
     2821 cactataggg cgaattgggg atcgatccac tagttctagc tagagcggcc gcataggcca
     2881 ctgtaagatt ccgccacatt ttatacactc tggtccttta actggcaaac cttcgggcgt
     2941 aatgcccaat ttttcgcctt tgtcttttgc ctttttcact tcacgtgctt ctggtacata
     3001 cttgcaattt atacagtgat gaccgctgaa tttgtatctt ccatagcatc tagcacatac
     3061 tcgattttta ccactccaat ctttataaaa atacttgatt ccctttctgg gacaagcaac
     3121 acagtgtttt agattctttt tttgtgatat tttatgctgt tctcccacac agcagcctcg
     3181 acatgatttc acttctattt tgttgccaag caagaaattt ttatggcctt ctatcgtaag
     3241 cccatataca gtactctcac cctggaaatc atccgtgaag ctgaaatata cgggttccct
     3301 ttttataatt ggcggaactt ctcttgtttt gtgaccactt cgacaatatg acaaaacatt
     3361 ctgtgaagtt gttcccccag gaattaattc gagctcgttt tcgacactgg atggcggcgt
     3421 tagtatcgaa tcgacagcag tatagcgacc agcattcaca tacgattgac gcatgatatt
     3481 actttctgcg cacttaactt cgcatctggg cagatgatgt cgaggcgaaa aaaaatataa
     3541 atcacgctaa catttgatta aaatagaaca actacaatat aaaaaaacta tacaaatgac
     3601 aagttcttga aaacaagaat ctttttattg tcagtactga ttagaaaaac tcatcgagca
     3661 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     3721 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     3781 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     3841 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     3901 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     3961 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     4021 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     4081 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     4141 ctgttttgcc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     4201 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     4261 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     4321 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     4381 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgaaacg tgagtctttt
     4441 ccttacccat ggttgtttat gttcggatgt gatgtgagaa ctgtatccta gcaagatttt
     4501 aaaaggaagt atatgaaaga agaacctcag tggcaaatcc taacctttta tatttctcta
     4561 caggggcgcg gcgtggggac aattcaacgc gtctgtgagg ggagcgtttc cctgctcgca
     4621 ggtctgcagc gaggagccgt aatttttgct tcgcgccgtg cggccatcaa aatgtatgga
     4681 tgcaaatgat tatacatggg gatgtatggg ctaaatgtac gggcgacagt cacatcatgc
     4741 ccctgagctg cgcacgtcaa gactgtcaag gagggtattc tgggcctcca tgtcgctggc
     4801 cgggtgaccc ggcggggacg aggcaagcta aacagatctg gcgcgcctta attaacccgc
     4861 agctggaatt ccacaccata gcttcaaaat gtttctactc cttttttact cttccagatt
     4921 ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt
     4981 tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa
     5041 aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt
     5101 ttctttttct tgaaaatttt ttttttgatt tttttctctt tcgatgacct cccattgata
     5161 tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt
     5221 acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagtttcc
     5281 taggttatta aatcattatc aatactcgcc atttcaaaga atacgtaaat aattaatagt
     5341 agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt
     5401 acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt gtctgggtga
     5461 acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc tggcatccag
     5521 aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt
     5581 ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac
     5641 ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc
     5701 atgtatctat ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa
     5761 agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt
     5821 atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat
     5881 tctactttta tagttagtct tttttttagt tttaaaacac cagaacttag tttcgatttc
     5941 ctaggttatt aaaatgaggc aggtttggtt ctcttggatt gtgggattgt tcctatgttt
     6001 tttcaacgtg tcttctgctg ctccagtcaa cactacaaca gaagatgaaa cggcacaaat
     6061 tccggctgaa gctgtcatcg gttactcaga tttagaaggg gatttcgatg ttgctgtttt
     6121 gccattttcc aacagcacaa ataacgggtt attgtttata aatactacta ttgccagcat
     6181 tgctgctaaa gaagaagggg tatctctcga gaaaagagag gctgaagctt ctaaaggtga
     6241 agaattattc actggtgttg tcccaatttt ggttgaatta gatggtgatg ttaatggtca
     6301 caaattttct gtctccggtg aaggtgaagg tgatgctact tacggtaaat tgaccttaaa
     6361 atttatttgt actactggta aattgccagt tccatggcca accttagtca ctactttcgg
     6421 ttatggtgtt caatgttttg ctagataccc agatcatatg aaacaacatg actttttcaa
     6481 gtctgccatg ccagaaggtt atgttcaaga aagaactatt tttttcaaag atgacggtaa
     6541 ctacaagacc agagctgaag tcaagtttga aggtgatacc ttagttaata gaatcgaatt
     6601 aaaaggtatt gattttaaag aagatggtaa cattttaggt cacaaattgg aatacaacta
     6661 taactctcac aatgtttaca tcatggctga caaacaaaag aatggtatca aagttaactt
     6721 caaaattaga cacaacattg aagatggttc tgttcaatta gctgaccatt atcaacaaaa
     6781 tactccaatt ggtgatggtc cagtcttgtt accagacaac cattacttat ccactcaatc
     6841 tgccttatcc aaagatccaa acgaaaagag agaccacatg gtcttgttag aatttgttac
     6901 tgctgctggt attacccatg gtatggatga attgtacaaa taaggtaccg cggctagcta
     6961 agatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     7021 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     7081 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
     7141 ggacgctcga agatccagct cagcgtacga cgccatttta agtccaaagg cacaatttta
     7201 cgttggaact aaacggtaaa gatcaaatat cacctttatt cttttttggt atgcatcgat
     7261 ttctttgcca gtaagaacta cgtgcaactc tacattagat caaaatataa cggatcataa
     7321 atgcaaacaa catttgtcca aaagaaacaa aaatgttggc gtttgtctcg aacatgacac
     7381 aatactcctt gatcgaggga cattgcccga ttaactatga tttatacttt ttactacagg
     7441 tactcgttct ccctactatt tcaccagtat acggtcgttt tatgattcat ttattgccta
     7501 taatggtaaa aagggacatc ttaaccttac gtgttgcaat gagaccattc ttttcgattc
     7561 atcaactgtg aaatttctga aatccagtac aatgcgaaac gctaccgata atggcaccgt
     7621 cttttgaatt atgaggccca gatatggcgt tcaatcttca acaaaaaaat gaatatcaaa
     7681 actaaattct atctggataa caatctttct ttggtttgca gaaatatcat taaaaaacaa
     7741 tccataacta ctaaggaagg gtataaatgg tttggtttac taattttata ttttgatttg
     7801 attagcaatt ataatctcta atatactatt tctgtagcca aaaatctgca atgtcaataa
     7861 gtagcaagtg ccgcttactc gtgagaatat caaccttata gcatatgttt tctatttttt
     7921 tttttttgct tatggagata atgaacattg tacacatgaa caaagtggta gttctctctc
     7981 cttgattttt tgtgcttgcg atatatataa ttgttacatt caatggccat gctaaataaa
     8041 ggttttagct ttgtttcact cgtgcccagg ataattaagc ttcagctggc ggccgccacg
     8101 gcgatatcgg atctcgaggc ctcggactag tggcgtaatc atggtcatag ctgtttcctg
     8161 tgtgaaattg ttatccgctc acaattccac acaacatacg agccgcggaa gcataaagtg
     8221 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     8281 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     8341 gagaggcggt ttgcgt
//
