LOCUS       62056_16625        7735 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 7735)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        285..388
                     /label=AmpR promoter
     CDS             389..1246
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1420..2008
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2194..2336)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2441..2629)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3404..3425)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3441..4520)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4521..4598)
                     /label=lacI promoter
     promoter        4911..4929
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4930..4954
                     /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             4969..4991
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5002..5019
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             5038..5691
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             5719..5739
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQS"
     CDS             7579..7596
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      7663..7710
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      361 tgcttcaata atttgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      421 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      481 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      541 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      601 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
      661 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      721 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      781 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      841 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      901 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      961 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1021 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1081 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1141 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1201 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1261 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1321 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1381 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1441 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1501 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1561 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1621 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1681 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1741 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1801 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1861 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1921 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1981 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2041 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2101 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2161 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2221 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2281 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2341 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2401 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2461 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2521 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2581 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2641 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2701 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2761 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2821 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2881 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2941 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     3001 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     3061 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     3121 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     3181 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     3241 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     3301 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     3361 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     3421 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3481 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     3541 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     3601 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     3661 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     3721 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     3781 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     3841 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     3901 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     3961 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4021 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4081 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     4141 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     4201 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     4261 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     4321 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     4381 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     4441 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     4501 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     4561 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     4621 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     4681 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     4741 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     4801 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     4861 cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact
     4921 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     4981 taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt agatctgatg
     5041 tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg
     5101 gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg
     5161 cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt
     5221 gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg
     5281 ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat
     5341 attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat
     5401 tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca
     5461 tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt
     5521 gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa
     5581 cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg
     5641 cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatgat
     5701 tacccagatc tgggtaccga gaacctgtac ttccaatcca atgatgggca tcaggacagc
     5761 ttacaggcgc tggcccccgc cagctgtcta gaaaacagct cccttgagca cacagtccat
     5821 agagagaaaa ctggaaaagg actaagtgct acgagactga gtgccagctc ggaggacatt
     5881 tctgacagac tggccggcgt ctctgtagga cttcccagct caacaacaac agaacaacca
     5941 aagccagcgg ttcaaacaaa aggcagaccc cacagtcagt gtttgaactc ctcccctttg
     6001 tctcatgctc aattaatgtt cccagcacca tcagcccctt gttcctctgc cccgtctgtc
     6061 ccagatattt ctaagcacag accccaggca tttgttccct gcaaaatacc ttccgcatct
     6121 cctcagacac agcgcaagtt ctctctacaa ttccagagga actgctctga acaccgagac
     6181 tcagaccagc tctccccagt cttcactcag tcaagacccc caccctccag taacatacac
     6241 aggccaaagc catcccgacc cgttccgggc agtacaagca aactagggga cgccacaaaa
     6301 agtagcatga cacttgatct gggcagtgct tccaggtgtg acgacagctt tggcggcggc
     6361 ggcaacagtg gcaacgccgt catacccagc gacgagacag tgttcacgcc ggtggaggac
     6421 aagtgcaggt tagatgtgaa caccgagctc aactccagca tcgaggacct tcttgaagca
     6481 tccatgcctt caagtgacac gacagtcact ttcaagtccg aagtcnatga agntgagaga
     6541 agtagcttta ncgatcgcca tgngantcag cgtctcagat gccctcccca tcgtcctcag
     6601 ntgcagtgga aaatggagaa gatattatca tcattcagca ggacacacca gaaactntna
     6661 ggacatacca aagcgaaaca gccttacaga gaagacgntg agtggctgaa aggccagcag
     6721 atagncctcg gagcattttc ttcctgttac caagcacagg atgtggggac tgggacttta
     6781 atggctgtga aacaggtgac gtacgtcaga aacacatcnt ccgagcagga ggaggtggtg
     6841 gaagcgttga gggaagagat ccggatgatg ggtcacctca accatccaaa catcatccgg
     6901 atgctggggg ccacgtgcga gaagagcaac tacaacctct tcattgagtg gatggcggga
     6961 ggatctgtgg ctcacctctt gagtaaatac ggagctttca aggagtcagt cgtcattaac
     7021 tacactgagc agttactgcg tggcctttcc tatctccacg agaaccagat cattcacaga
     7081 gacgtcaaag gtgccaacct gctcattgac agcaccggtc agaggctgag aattgcagac
     7141 tttggagctg ctgccaggtt ggcatcaaaa ggaaccggtg caggagagtt ccagggacag
     7201 ttactgggga caattgcatt catggcgcct gaggtcctaa gaggtcagca gtatggtagg
     7261 agctgtgatg tatggagtgt tggctgcgcc attatagaaa tggcttgtgc aaaaccacct
     7321 tggaatgcag aaaaacactc caatcatctc gccttgatat ttaagattgc tagcgcaact
     7381 actgcaccgt ccatcccgtc acacctgtcc ccgggtctgc gcgacgtggc gctgcgctgc
     7441 ttagaacttc agcctcagga ccggcctccg tccagagagc tgctgaaaca tccggtcttc
     7501 cgtaccacgt ggtagcattg gaagtggata acggatccga attcgagctc cgtcgacaag
     7561 cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     7621 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     7681 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggat
//
