LOCUS       62056_9610        5977 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 5977)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      489..536
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(711..739)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        852..956
                     /label=AmpR promoter
     CDS             957..1814
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1988..2576
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2762..2904)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3009..3197)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(4475..4496)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4512..5591)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5592..5669)
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggccgataga caaggaactg tatcctttag
      121 cttccctcag atcactcttt ggcagcgacc cctcgtcaca ataaagatag gggggcaatt
      181 aaaggaagct ctattagata caggagcaga tgatacagta ttagaagaaa tgaatttgcc
      241 aggaagatgg aaaccaaaaa tgataggggg aattggaggt tttatcaaag taagacagta
      301 tgatcagata ctcatagaaa tctgcggaca taaagctata ggtacagtat tagtaggacc
      361 tacacctgtc aacataattg gaagaaatct gttgactcag attggttgca ctttaaattt
      421 ttagggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      481 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      541 aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
      601 tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt
      661 catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt
      721 atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc gtgatacgcc
      781 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
      841 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      901 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      961 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1021 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1081 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1141 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1201 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1261 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1321 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1381 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1441 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1501 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1561 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1621 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1681 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1741 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1801 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1861 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1921 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1981 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2041 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2101 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2161 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2221 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2281 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2341 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2401 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2461 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2521 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2581 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2641 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2701 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2761 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatggtg
     2821 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     2881 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     2941 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3001 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     3061 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     3121 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     3181 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     3241 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     3301 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     3361 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     3421 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     3481 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     3541 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     3601 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     3661 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
     3721 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     3781 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     3841 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     3901 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     3961 tgacgatcag cggtccagtg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     4021 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     4081 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     4141 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     4201 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     4261 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     4321 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     4381 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     4441 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     4501 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4561 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     4621 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4681 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     4741 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     4801 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     4861 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     4921 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     4981 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     5041 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     5101 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     5161 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     5221 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     5281 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     5341 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     5401 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     5461 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     5521 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     5581 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     5641 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     5701 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     5761 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     5821 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     5881 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     5941 tccggcgtag aggatcgaga tctcgatccc gcgaaat
//
