LOCUS       62056_11510        5329 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 5329)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    359..380
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        395..425
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    433..449
                     /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)."
     CDS             469..642
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     promoter        1098..1126
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             1173..1826
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             1833..1856
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     CDS             1884..2225
                     /codon_start=1
                     /label=GFP nanobody
                     /note="GFP-binding fragment of a single-chain camelid
                     antibody (Rothbauer et al., 2008)"
                     /translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
                     WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCNVNVGFEYW
                     GQGTQVTVSS"
     promoter        2547..2651
                     /label=AmpR promoter
     CDS             2652..3509
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3683..4271
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4515..4592
                     /label=lacI promoter
     CDS             join(4593..5329,1..343)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
       61 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
      121 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
      181 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
      241 aaaaaccacg ctagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      301 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      361 atgtaagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
      421 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
      481 acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      541 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      601 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt
      661 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact
      721 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgtg
      781 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
      841 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
      901 gatggcgttg gaattagctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag
      961 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
     1021 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
     1081 tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
     1141 cggataacaa tttcacacag gaaacagtat tcatgtcccc tatactaggt tattggaaaa
     1201 ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag
     1261 agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt
     1321 tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg
     1381 ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg
     1441 cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg
     1501 catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc
     1561 tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc
     1621 atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc
     1681 tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg
     1741 ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt
     1801 ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag gggcccctgg
     1861 gatccccgga attcatggcg caggttcagc tggttgaaag cggtggtgca ctggttcagc
     1921 ctggtggtag cctgcgtctg agctgtgcag caagcggttt tccggttaat cgttatagca
     1981 tgcgttggta tcgtcaggca ccgggtaaag aacgtgaatg ggttgcaggt atgagcagtg
     2041 ccggtgatcg tagcagctat gaagatagcg ttaaaggtcg ttttaccatc agccgtgatg
     2101 atgcacgtaa taccgtttat ctgcaaatga atagcctgaa accggaagat accgcagtgt
     2161 attattgcaa tgttaacgtg ggctttgaat attggggtca gggcacccag gttaccgtta
     2221 gcagcaaact cgagcggccg catcgtgact gactgacgat ctgcctcgcg cgtttcggtg
     2281 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2341 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2401 gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga agacgaaagg
     2461 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     2521 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2581 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2641 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2701 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2761 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2821 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2881 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2941 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3001 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3061 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3121 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3181 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3241 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3301 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3361 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3421 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3481 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3541 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3601 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3661 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3721 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3781 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3841 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3901 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3961 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4021 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4081 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4141 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4201 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4261 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4321 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4381 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4441 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4501 accgcataaa ttccgacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     4561 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     4621 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     4681 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     4741 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     4801 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     4861 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     4921 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     4981 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     5041 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     5101 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     5161 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     5221 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     5281 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaa
//
