LOCUS       62056_11485        5719 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 5719)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             918..935
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             969..1679
                     /codon_start=1
                     /label=TagRFP
                     /note="monomeric derivative of red fluorescent protein from
                     Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             1686..1703
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     promoter        2022..2126
                     /label=AmpR promoter
     CDS             2127..2984
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3158..3746
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3990..4067
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4068..5147
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    5163..5184
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5199..5229
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             5273..5446
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatccccggg aatttccggt ggtggtggtg
      961 gaattctaat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt
     1021 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
     1081 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
     1141 ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc aaccacaccc
     1201 agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca
     1261 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     1321 gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc
     1381 agaagaaaac actcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc
     1441 tggaaggcag aagcgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact
     1501 tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
     1561 atgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
     1621 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac aaacttaatc
     1681 tcgagcacca ccaccaccac cactagaagc ttattcatcg tgactgactg acgatctgcc
     1741 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     1801 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     1861 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt
     1921 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1981 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2041 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2101 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2161 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2221 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2281 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2341 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     2401 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2461 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2521 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2581 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2641 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2701 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2761 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2821 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2881 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2941 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3001 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3061 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3121 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3181 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3241 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3301 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3361 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3421 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3481 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3541 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3601 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3661 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3721 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3781 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3841 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3901 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3961 tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggtgcaaa acctttcgcg
     4021 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     4081 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     4141 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     4201 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     4261 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     4321 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     4381 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     4441 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     4501 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     4561 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     4621 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     4681 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     4741 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     4801 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     4861 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     4921 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     4981 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     5041 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5101 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5161 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     5221 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     5281 gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5341 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5401 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg
     5461 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga
     5521 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
     5581 cgtaacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
     5641 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
     5701 ttttgatggc gttggaatt
//
