LOCUS       62056_10230        5373 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 5373)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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)."
     RBS             83..103
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             123..140
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      207..254
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      291..746
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        773..877
                     /label=AmpR promoter
     CDS             878..1735
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1909..2497
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2683..2822)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2927..3115)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3890..3911)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3927..5006)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5007..5084)
                     /label=lacI promoter
ORIGIN
        1 gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagtaataa ttttgtttaa ctttaagaag gaggtctaga atgcatctcg
      121 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
      181 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      241 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
      301 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
      361 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
      421 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
      481 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
      541 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
      601 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
      661 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
      721 taacaaaata ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac
      781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      841 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      901 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      961 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1021 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1081 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1141 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1201 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1261 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1321 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1381 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1441 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1501 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1561 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1621 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1681 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1741 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1801 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1861 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1921 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1981 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2041 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2101 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2161 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2221 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2281 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2341 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2401 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2461 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2521 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2581 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2641 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2701 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2761 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2821 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2881 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2941 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3001 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3061 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3121 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3181 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3241 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3301 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3361 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3421 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3481 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3541 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     3601 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3661 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3721 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     3781 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     3841 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     3901 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3961 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4021 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4081 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4141 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4201 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4261 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4321 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4381 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4441 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4501 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4561 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4621 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4681 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4741 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     4801 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     4861 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     4921 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     4981 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5041 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5101 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5161 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5221 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5281 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5341 gccggccacg atgcgtccgg cgtagaggat cga
//
