LOCUS       Exported                6996 bp ds-DNA     circular SYN 17-MAR-2017
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pet151DTOPOBrox1-411 K141E
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6996)
  AUTHORS   Path-IT
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Mar 17, 2017 from SnapGene 3.3.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6996
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          267..289
                     /organism="Enterobacteria phage T7"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10760"
     promoter        209..227
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             300..317
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             318..359
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /note="V5 tag"
                     /translation="GKPIPNPLLGLDST"
     CDS             360..380
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /note="TEV site"
                     /translation="ENLYFQG"
     terminator      1708..1755
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2079..2183
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             2184..3044
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3215..3803
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3989..4131
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(4233..4424)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5736..6818)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6819..6896)
                     /gene="lacI"
                     /note="lacI promoter"
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgc
      301 atcatcacca tcaccatggt aagcctatcc ctaaccctct cctcggtctc gattctacgg
      361 aaaacctgta ttttcaggga attgatccct tcaccatgac ccattggttt cataggaacc
      421 cattaaaagc cacagctcct gtgtctttta attactatgg tgtagtcact ggcccttctg
      481 cttcaaaaat atgcaatgac ttgaggtcat ccagggcacg actccttgaa ctgttcactg
      541 atttgagctg taatccagaa atgatgaaga atgcagcaga ttcatatttc tcacttttac
      601 aaggtttcat aaattctttg gatgaatcta cccaagaaag caagttacga tatattcaaa
      661 atttcaagtg gactgataca ttgcaaggac aggttccaag tgcccagcag gatgctgttt
      721 ttgaattaat ttccatggga tttaatgtag ctttatggta taccaaatat gcttcaagac
      781 tggctggaaa agaaaatata acagaagatg aagcagaaga agttcatcga agcctaaaga
      841 ttgcagctgg gatttttaaa catttaaagg aaagtcatct cccaaaactc attacacctg
      901 cggaaaaagg aagagattta gagtcacgac tcatagaagc atacgttatt caatgtcagg
      961 ctgaagctca agaagtaaca attgctcgag caattgaact aaaacatgct cctggactaa
     1021 ttgctgcact ggcgtatgaa acagccaatt tctatcaaaa agctgatcat actttatcca
     1081 gtttggagcc tgcatattct gccaaatgga gaaaatatct tcacttgaag atgtgtttct
     1141 acacagctta tgcttactgt taccatggtg agactttatt ggctagtgat aaatgcggtg
     1201 aagcaatcag gtctctccaa gaagcagaaa aattgtatgc aaaggcagaa gcactgtgta
     1261 aagaatatgg agaaaccaaa ggacctggac caacagtcaa accttcagga catctgttct
     1321 ttaggaaact tggaaacctt gtgaagaaca ccctagaaaa atgtcagaga gaaaatggat
     1381 ttatttactt tcaaaaaact ccaacagaag ccccacagct ggaactcaaa gcaaattatg
     1441 gtctcgtaga gcctatacct ttcgaatttc ctcctacaag tgttcagtgg acaccagaaa
     1501 cattggctgc atttgatctc accaaaagac ccaaggatga cagtactaaa cccaaaccag
     1561 aagaagaagt gaaacctgtg aaagaaccag acatcaaacc tcaaaaggac actgggtgct
     1621 acatctccta aaagggcgag ctcagatccg gctgctaaca aagcccgaaa ggaagctgag
     1681 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     1741 ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca
     1801 gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga
     1861 tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata
     1921 aactaccgca ttaaagctag cttatcgatg ataagctgtc aaacatgaga attaattctt
     1981 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     2041 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2101 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2161 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2221 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2281 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2341 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2401 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     2461 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2521 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2581 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2641 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2701 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     2761 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2821 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2881 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2941 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3001 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3061 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3121 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3181 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3241 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3301 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3361 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3421 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3481 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3541 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3601 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3661 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3721 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3781 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3841 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3901 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3961 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4021 gcggtatttc acaccgcata tatggtgcac tctcagtaca atctgctctg atgccgcata
     4081 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4141 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4201 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4261 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4321 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4381 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4441 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4501 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4561 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4621 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4681 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4741 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4801 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4861 aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc
     4921 cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc
     4981 gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta
     5041 gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca
     5101 acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg
     5161 tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccagtgatc gaagttaggc
     5221 tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg
     5281 acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg
     5341 ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg
     5401 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     5461 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     5521 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     5581 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     5641 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     5701 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5761 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5821 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     5881 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     5941 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     6001 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     6061 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     6121 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     6181 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     6241 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     6301 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     6361 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     6421 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     6481 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     6541 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     6601 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     6661 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     6721 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     6781 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     6841 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     6901 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     6961 gccgttgagc accgccgccg caaggaatgg tgcatg
//
