LOCUS       62056_9615        6181 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 6181)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6181
                     /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)"
     CDS             98..115
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      693..740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(915..943)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        1056..1160
                     /label=AmpR promoter
     CDS             1161..2018
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2192..2780
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2966..3108)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3213..3401)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(4679..4700)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4716..5795)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5796..5873)
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggctagccat caccaccatc accatggcgc
      121 gagcaaagga gaagaactct tcactggagt tgtcccaatt cttgttgaat tagatggtga
      181 tgttaacggc cacaagttct ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      241 acttaccctg aagttcatct gcactactgg caaactgcct gttccatggc caacactagt
      301 cactactctg tgctatggtg ttcaatgctt ttcaagatac ccggatcata tgaaacggca
      361 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaaggacca tcttcttcaa
      421 agatgacggc aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa
      481 tagaatcgag ttaaaaggta ttgacttcaa ggaagatggc aacattctgg gacacaaatt
      541 ggaatacaac tataactcac acaacgttct catcatggca gacaaacaag gtggctctgg
      601 ctctggctcg agcaatcccg ggaattaagg atccggctgc taacaaagcc cgaaaggaag
      661 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      721 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
      781 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
      841 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
      901 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
      961 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1021 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1081 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1141 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1201 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1261 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1321 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1381 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     1441 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1501 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1561 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1621 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1681 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     1741 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1801 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1861 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1921 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1981 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2041 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2101 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2161 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2221 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2281 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2341 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2401 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2461 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2521 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2581 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2641 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2701 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2761 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2821 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2881 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2941 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3001 gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3061 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     3121 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3181 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3241 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     3301 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     3361 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     3421 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     3481 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     3541 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     3601 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     3661 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     3721 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     3781 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     3841 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc
     3901 gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca
     3961 ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg
     4021 aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg
     4081 cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc
     4141 tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc agtgatcgaa gttaggctgg
     4201 taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca
     4261 gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga
     4321 aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca
     4381 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     4441 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     4501 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     4561 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     4621 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     4681 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4741 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4801 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     4861 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     4921 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     4981 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     5041 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     5101 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     5161 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     5221 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     5281 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     5341 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     5401 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     5461 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     5521 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     5581 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     5641 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     5701 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     5761 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     5821 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     5881 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     5941 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     6001 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     6061 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6121 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
     6181 t
//
