LOCUS       62056_22915        8099 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 8099)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      69..1411
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1438..1542
                     /label=AmpR promoter
     CDS             1543..2400
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2574..3162
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3450..3471
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3486..3516
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3524..3540
                     /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     3548..3564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3585..3603
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(3643..3807)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(4458..4721)
                     /codon_start=1
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAE"
     promoter        complement(5162..5180)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5200..5843
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     5849..5865
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5899..5915)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5918..6165
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(6184..6202)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6212..6228)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6373..6828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        complement(7358..7765)
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
      121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
      181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
      241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
      301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
      361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
      421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
      481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
      541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
      601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
      661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
      721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
      781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
      841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
      901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
      961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
     1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
     1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
     1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
     1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
     1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
     1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
     1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
     3481 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
     3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     3601 agggaacaaa agctggagct cgctattacg ccagctgaat tggagcgacc tcatgctata
     3661 ctgagaaagc aacctgacct acaggaaaga gttactcaag aataagaatt ttcgttttaa
     3721 aacctaagag tcactttaaa atttgtatac acttattttt tttataactt atttaataat
     3781 aaaaatcata aatcataaga aattcgctta tttagaagtg tcaacaacgt atctaccaac
     3841 gatttgaccc ttttccatct tttcgtaaat ttctggcaag gtagacaagc cgacaacctt
     3901 gattggagac ttgaccaaac ctctggcgaa gaattgttaa ttaaagatct ccgcggttac
     3961 tctttctttc ttaatctgtc ttgaagattt ctagaggaag agaaactacg cataacttct
     4021 gctcttacta tttcccatgc acatggagaa tatgattttt cctgtagata taaggttaat
     4081 ctatgaaaat atttacgcac agccaatata gaatcttctt ctagtaatgg agtaccttct
     4141 aaacccgctt cttgcataac gcaagcttct agatctctta gctgttggtc caatccagta
     4201 cagaattgat gcaataatga ttcgttccat gcggcggctg aaccttctgt ggaaaacaat
     4261 tgaaatgtct gctgtaacat ttcgtgaact aaggccatag cctgagcttt ctgcacttga
     4321 ttaccgccaa aagcttcatg aggagatccg aaatctcttc tatgatccaa gcaagagaag
     4381 ggtgagattc tacgcatttg agcaagcaga cgcaaggctc ttgtatgagc tagtgaatgt
     4441 gtttgtggca agtcacattc agcctctctc ttctcgagag ataccccttc ttctttagca
     4501 gcaatgctgg caatagtagt atttataaac aataacccgt tatttgtgct gttggaaaat
     4561 ggcaaaacag caacatcgaa atccccttct aaatctgagt aaccgatgac agcttcagcc
     4621 ggaatttgtg ccgtttcatc ttctgttgta gtgttgactg gagcagctaa tgcggaggat
     4681 gctgcgaata aaacagcagt aaaaattgaa ggaaatctca tagcggccgc gctagttcta
     4741 gaaaacttag attagattgc tatgctttct ttctaatgag caagaagtaa aaaaagttgt
     4801 aatagaacaa gaaaaatgaa actgaaactt gagaaattga agaccgttta ttaacttaaa
     4861 tatcaatggg aggtcatcga aagagaaaaa aatcaaaaaa aaaattttca agaaaaagaa
     4921 acgtgataaa aatttttatt gcctttttcg acgaagaaaa agaaacgagg cggtctcttt
     4981 tttcttttcc aaacctttag tacgggtaat taacgacacc ctagaggaag aaagagggga
     5041 aatttagtat gctgtgcttg ggtgttttga agtggtacgg cgatgcgcgg agtccgagaa
     5101 aatctggaag agtaaaaaag gagtagaaac attttgaagc tatgagctcc agcttttgtt
     5161 ccctttagtg agggttaatt gcgcgccgag ctcagtttat cattatcaat actcgccatt
     5221 tcaaagaata cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat
     5281 tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa
     5341 tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact aaatataatg
     5401 gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt
     5461 ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg
     5521 cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa
     5581 tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac accttctatt
     5641 accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga
     5701 aattattccc ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg
     5761 taaatctatt tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt
     5821 aaaacaccag aacttagttt cgacggattc tagaactagt ggatcccccg ggctgcagga
     5881 attcgatatc aagcttatcg ataccgtcga cctcgagtca tgtaattagt tatgtcacgc
     5941 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     6001 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     6061 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     6121 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccaatt
     6181 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     6241 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     6301 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     6361 gcgaatggcg cgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6421 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6481 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6541 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6601 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6661 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6721 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6781 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttcc tgatgcggta
     6841 ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt
     6901 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttc
     6961 atgagatgcc tgcaagcaat tcgttctgta tcaggcgcag gagcgtcccg tccgggtcga
     7021 ccaaagcggc catcgtgcct ccccactcct gcagttcggg ggcatggatg cgcggatagc
     7081 cgctgctggt ttcctggatg ccgacggatt tgcactgccg gtagaactcc gcgaggtcgt
     7141 ccagcctcag gcagcagctg aaccaactcg cgaggggatc gagcccgggg tgggcgaaga
     7201 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     7261 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg
     7321 gcgtcgcttg gtcggtcatt tcgaattcga gctcgccctt agattagatt gctatgcttt
     7381 ctttctaatg agcaagaagt aaaaaaagtt gtaatagaac aagaaaaatg aaactgaaac
     7441 ttgagaaatt gaagaccgtt tattaactta aatatcaatg ggaggtcatc gaaagagaaa
     7501 aaaatcaaaa aaaaaaattt tcaagaaaaa gaaacgtgat aaaaattttt attgcctttt
     7561 tcgacgaaga aaaagaaacg aggcggtctc ttttttcttt tccaaacctt tagtacgggt
     7621 aattaacgac accctagagg aagaaagagg ggaaatttag tatgctgtgc ttgggtgttt
     7681 tgaagtggta cggcgatgcg cggagtccga gaaaatctgg aagagtaaaa aaggagtaga
     7741 aacattttga agctatggtg tgtggcatat gcgtatatat accaatctaa gtctgtgctc
     7801 cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg
     7861 aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg
     7921 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     7981 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     8041 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
