LOCUS       62056_19975        6885 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 6885)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      325..1667
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(1725..2396)
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(2520..2536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2544..2560)
                     /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)."
     promoter        complement(2568..2598)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2613..2634)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2922..3510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3684..4541)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(4745..5767)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     6302..6318
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6328..6346
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6862..6878)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catcattcca ctcaacattc aggctcctct gcgcacgtaa agtccaaagg caataccctg
       61 ctcggtggaa tgcgccgggc ttgtcgattt tacgcacata tgcgcattct tgacttgaag
      121 cggaggagtt cttcgttgcg ggttacagtg ttttaataaa agaatggtca aatcaaactg
      181 ctagatatac ctgtcagaca ctctagttgt tgacccctat actcttaata catcagacag
      241 tacatgcatg ttgcatgatg atgataatgt ctgtttagat tccaagtgtc tactgctggc
      301 ctagtttctc ggtactatgc atatgatcca atatcaaagg aaatgatagc attgaaggat
      361 gagactaatc caattgagga gtggcagcat atagaacagc taaagggtag tgctgaagga
      421 agcatacgat accccgcatg gaatgggata atatcacagg aggtactaga ctacctttca
      481 tcctacataa atagacgcat ataagtacgc atttaagcat aaacacgcac tatgccgttc
      541 ttctcatgta tatatatata caggcaacac gcagatatag gtgcgacgtg aacagtgagc
      601 tgtatgtgcg cagctcgcgt tgcattttcg gaagcgctcg ttttcggaaa cgctttgaag
      661 ttcctattcc gaagttccta ttctctagaa agtataggaa cttcagagcg cttttgaaaa
      721 ccaaaagcgc tctgaagacg cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc
      781 tactaaaata ttgcgaatac cgcttccaca aacattgctc aaaagtatct ctttgctata
      841 tatctctgtg ctatatccct atataaccta cccatccacc tttcgctcct tgaacttgca
      901 tctaaactcg acctctacat tttttatgtt tatctctagt attactcttt agacaaaaaa
      961 attgtagtaa gaactattca tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc
     1021 tatacgtagt atatagagac aaaatagaag aaaccgttca taattttctg accaatgaag
     1081 aatcatcaac gctatcactt tctgttcaca aagtatgcgc aatccacatc ggtatagaat
     1141 ataatcgggg atgcctttat cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa
     1201 acgcgggaag tggagtcagg ctttttttat ggaagagaaa atagacacca aagtagcctt
     1261 cttctaacct taacggacct acagtgcaaa aagttatcaa gagactgcat tatagagcgc
     1321 acaaaggaga aaaaaagtaa tctaagatgc tttgttagaa aaatagcgct ctcgggatgc
     1381 atttttgtag aacaaaaaag aagtatagat tctttgttgg taaaatagcg ctctcgcgtt
     1441 gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc
     1501 gttgcatttt tgttttacaa aaatgaagca cagattcttc gttggtaaaa tagcgctttc
     1561 gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct
     1621 ctcgcgttgc atttttgttc tacaaaatga agcacagatg cttcgttggg ctgcaggcaa
     1681 gtgcacaaac aatacttaaa taaatactac tcagtaataa cctatttctt agcatttttg
     1741 acgaaatttg ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca
     1801 tcaacaccaa taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca
     1861 ccagctaaca taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc
     1921 gagttccaat ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac
     1981 gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt
     2041 cttttaataa ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc
     2101 agttggacga tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga
     2161 ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca
     2221 agacttgaaa ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat
     2281 ataataccca gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc
     2341 aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatactc
     2401 caagctgcct ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca ccaatgccct
     2461 ccctcttggc cctctccttt tctttagtga gggttaattg cgcgcttggc gtaatcatgg
     2521 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2581 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2641 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2701 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2761 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2821 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3661 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3781 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3841 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3961 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaattc
     4561 aatgcgtttt attcttgttg acatggagct attaaatcac tagaaggcac tctttgctgc
     4621 ttggacaaat gaacgtatct tatcgagatc ctgaacacca tttgtctcaa ctccggagct
     4681 gacatcgaca ccaacgatct tatatccaga ttcgtcaagc tgtttgatga tttcagtaac
     4741 gctattcctt tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt
     4801 ctacacagcc atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag
     4861 tcccggctcc ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga
     4921 aattgccgtc aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc
     4981 ggagccgcgg cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct
     5041 ccatacaagc caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc
     5101 cgaacatcgc ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat
     5161 tgttggagcc gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa
     5221 gcatcagctc atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt
     5281 gccagtgata cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt
     5341 gaccgattcc ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag
     5401 cgatcgcatc catggcctcc gcgaccggct gcagaacagc gggcagttcg gtttcaggca
     5461 ggtcttgcaa cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctga
     5521 attccccaat gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa
     5581 cataacgatc tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc
     5641 ctcctacatc gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg
     5701 agacgctgtc gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct
     5761 ttttcatttt gaagattggg ttcctctttg agagagttaa tagacttgaa ttgaggaatg
     5821 gaaatcgggc tgagctcact gctttatatg tgagaaaagg tgatgagcag gtggtaagag
     5881 gcagattttt gtggtcggtg ctgtgttttg ttgggtttga gctaggtccg gtgagggagg
     5941 ggcaagctaa atagctacca gccctgagga cgtcacagag ttttagtagt tctttttctg
     6001 ccatcggaca tgcagctcac gcctagaccc aattacaccc tttgcgccct cccacacatc
     6061 cggccggttg gatcattgtc agatacggca gagaaatcgc aacctcggcc aagggtagcc
     6121 acgattcgaa gccgcggagt ctagattaac gcttacaatt tccattcgcc attcaggctg
     6181 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6241 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     6301 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtact
     6361 caaattggtt ataaatgtag gtattacctg tacattttat ttattcgaga aaaacaaaca
     6421 aaccaaaacg atgcagcgat cactaccggc gtcatccgga attgaaacgg cgcgcgttga
     6481 ggggggcctt cttgttcgga cacacacggg gccgccaaag tggggcccat acggctgccc
     6541 atcccttgct ctccaatacc ctggtcatgt gactcgaaat tggagactca caaagatgct
     6601 ttgaaacccc gccgggaact ttttctcaca gacacaatca gtcgacctta tcatcgcaaa
     6661 atcgacaaac ctcaaaaaga actcgaagag tcagtctccc ttgcggttaa actgattgat
     6721 atctcgtgtt ctctaaacct tcaataagtg agttactgcg cacaaagccg actatactga
     6781 gcaccaacgc gaattgctca cgttgtcgcc taacagatct tggctttcgt aggaacccaa
     6841 tcttcaaaat ggctcgaggg ggatccacta gttctagagc ggccg
//
