LOCUS       Exported                7400 bp ds-DNA     circular SYN 06-MAR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  AUTHORS   MReijns
  TITLE     Direct Submission
  JOURNAL   Exported Mar 6, 2018 from SnapGene Viewer 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7400
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    972..1907
                     /label=RNASEH2B
     RBS             1922..1944
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1952..2446
                     /codon_start=1
                     /label=RNASEH2C
                     /translation="MESGDEAAIERHRVHLRSATLRDAVPATLHLLPCEVAVDGPAPVG
                     RFFTPAIRQGPEGLEVSFRGRCLRGEEVAVPPGLVGYVMVTEEKKVSMGKPDPLRDSGT
                     DDQEEEPLERDFDRFIGATANFSRFTLWGLETIPGPDAKVRGALTWPSLAAAIHAQVPE
                     D"
     RBS             2454..2476
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2484..3383
                     /codon_start=1
                     /label=RNASEH2A
                     /translation="MDLSELERDNTGRCRLSSPVPAVCRKEPCVLGVDEAGRGPVLGPM
                     VYAICYCPLPRLADLEALKVADSKTLLESERERLFAKMEDTDFVGWALDVLSPNLISTS
                     MLGRVKYNLNSLSHDTATGLIQYALDQGVNVTQVFVDTVGMPETYQARLQQSFPGIEVT
                     VKAKADALYPVVSAASICAKVARDQAVKKWQFVEKLQDLDTDYGSGYPNDPKTKAWLKE
                     HVEPVFGFPQFVRFSWRTAQTILEKEAEDVIWEDSASENQEGLRKITSYFLNEGSQARP
                     RSSHRYFLERGLESATSL"
     promoter        3703..3807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3808..4668
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4839..5427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5671..5748
                     /gene="lacI (mutant)"
                     /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             5749..6831
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    6844..6865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6880..6910
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6918..6934
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     6942..6958
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6954..7400
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
     primer_bind     complement(6974..6990)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 gaagttctgt tccaggggcc cctgggatcc ccggaattcc
      961 cgggtcgact cgccgctggc gtggactgcg gggacggggt tggcgcccgg cagcacgtgt
     1021 tcctggtttc agaatattta aaagatgctt caaagaagat gaaaaatggg ctaatgtttg
     1081 taaaactggt taacccctgt tcaggagaag gagccattta cttgttcaat atgtgtctac
     1141 agcagctgtt tgaagtaaaa gttttcaagg aaaaacacca ttcttggttt ataaatcaat
     1201 cagttcaatc aggaggtctt ctccattttg ccacacctgt ggatcctcta tttctgcttc
     1261 tccactacct cataaaggct gataaggagg ggaagtttca gccccttgat caagttgtgg
     1321 tggataacgt gtttccaaat tgcatcttgt tgctgaaact tcctggactt gagaagttac
     1381 ttcatcatgt gacagaggaa aaaggtaatc cagaaataga caacaagaaa tattacaagt
     1441 acagcaaaga gaagacatta aagtggctgg aaaaaaaggt taatcaaact gtggcagcat
     1501 taaaaaccaa taatgtgaat gtcagttccc gggtacagtc aactgcattt ttctctggtg
     1561 accaagcttc cactgacaag gaagaggatt atattcgtta tgcccatggt ctgatatctg
     1621 actacatccc taaagaatta agtgatgact tatctaaata cttaaagctt ccagaacctt
     1681 cagcctcatt gccaaatcct ccatcaaaga aaataaagtt atcagatgag cctgtagaag
     1741 caaaagaaga ttacactaag tttaatacta aagatttgaa gactgaaaag aaaaatagca
     1801 aaatgactgc agctcagaag gctttggcta aagttgacaa gagtggaatg aaaagtattg
     1861 ataccttttt tggggtaaaa aataaaaaaa aaattggaaa ggtttgaaac gaccatcgat
     1921 ttttgtttaa ctttaagaag gagatataca tatggagagc ggcgacgaag cggccatcga
     1981 gaggcaccgc gtccacttgc gctccgccac attgcgcgac gccgtacccg ccacactgca
     2041 tctgctgccc tgcgaggttg cggtggacgg gcccgccccg gtggggcgct tcttcacgcc
     2101 cgccatccgc cagggccccg agggactcga agtgtcgttt cggggccgct gtctacgggg
     2161 agaggaggtg gcggtgccgc ctggcctcgt gggatacgtg atggtgacag aagagaagaa
     2221 ggtgtcgatg gggaagccag accccttgcg ggattccggg actgacgacc aagaggagga
     2281 gccgctggag cgggacttcg accgcttcat tggagccact gccaacttca gccgcttcac
     2341 cctgtggggt ctggagacca tccctggccc ggatgccaaa gtgcgtgggg ccttaacttg
     2401 gcccagcctt gcggcagcga ttcacgcaca ggtgcccgag gactagggta ccttttgttt
     2461 aactttaaga aggagatata catatggatc tcagcgagct ggagagagac aatacaggcc
     2521 gctgtcgcct gagttcgcct gtgcccgcgg tgtgccgcaa ggagccttgc gtcctgggcg
     2581 tcgatgaggc gggcaggggc cccgtgctgg gccccatggt ctacgccatc tgttattgtc
     2641 ccctgcctcg cctggcagat ctggaggcgc tgaaagtggc agactcaaag accctattgg
     2701 agagcgagcg ggaaaggctg tttgcgaaaa tggaggacac ggactttgtc ggctgggcgc
     2761 tggatgtgct gtctccaaac ctcatctcta ccagcatgct tgggcgggtc aaatacaacc
     2821 tgaactcctt gtcacatgat acagccactg ggcttataca gtatgcattg gaccagggcg
     2881 tgaacgtcac ccaggtattc gtggacaccg tagggatgcc agagacatac caggcgcggc
     2941 tgcagcaaag ttttcccggg attgaggtga cggtcaaggc caaagcagat gccctctacc
     3001 cggtggttag tgctgccagc atctgtgcca aggtggcccg ggaccaggcc gtgaagaaat
     3061 ggcagttcgt ggagaaactg caggacttgg atactgatta tggctcaggc taccccaatg
     3121 atcccaagac aaaagcgtgg ttgaaggagc acgtggagcc tgtgttcggc ttcccccagt
     3181 ttgtccggtt cagctggcgc acggcccaga ccatcctgga gaaagaggcg gaagatgtta
     3241 tatgggagga ctcagcatcc gagaatcagg agggactcag gaagatcaca tcctacttcc
     3301 tcaatgaagg gtcccaagcc cgtccccgtt cttcccaccg atatttcctg gaacgcggcc
     3361 tggagtcagc aaccagcctc taggagctcg cggccgcatc gtgactgact gacgatctgc
     3421 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3481 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3541 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat
     3601 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3661 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3721 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3781 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3841 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3901 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3961 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4021 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     4081 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4141 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4201 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4261 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4321 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4381 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4441 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4501 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4561 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4621 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4681 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4741 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4801 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4861 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4921 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4981 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5041 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5101 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5161 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5221 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5281 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5341 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5401 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5461 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5521 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5581 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     5641 ctgtgcggta tttcacaccg cataaattcc gacaccatcg aatggtgcaa aacctttcgc
     5701 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     5761 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     5821 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     5881 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     5941 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     6001 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     6061 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     6121 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     6181 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     6241 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     6301 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     6361 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     6421 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     6481 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     6541 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     6601 ccgtcaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     6661 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     6721 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6781 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6841 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6901 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6961 tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     7021 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     7081 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct
     7141 ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg
     7201 atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca
     7261 acgtaaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt
     7321 gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta
     7381 tttttgatgg cgttggaatt
//
