LOCUS       Exported                6746 bp DNA     circular SYN 08-OCT-2021
DEFINITION  pH6Sumo-SnRK2.6 (aa11-362) Circularized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6746
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..318
                     /label=His6-SUMO tag
                     /note="color: #cc0000; direction: RIGHT"
     CDS             7..24
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             28..318
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="color: #cc99b2
                     Cleavage site after base 318"
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     misc_feature    322..1380
                     /label=SnRK2.6 (aa11-362)
                     /note="color: #cc0000; direction: RIGHT"
     misc_feature    1381..1386
                     /label=destroyed XhoI site
                     /note="color: #cc0000; direction: RIGHT"
     CDS             1387..1404
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      1471..1518
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      1555..2010
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2037..2141
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2142..3002
                     /label=AmpR
                     /note="color: #cc0000; direction: RIGHT"
     CDS             2142..3002
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 2142 .. 2210 / #ccffcc / signal sequence
                        2: 2211 .. 3002 / #ccffcc
                     Cleavage site after base 2210"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3173..3761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    3947..4089
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(4191..4382)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5191..6273)
                     /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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     misc_feature    complement(5191..6150)
                     /label=LacI
                     /note="color: #cc0000; direction: LEFT"
     promoter        complement(6274..6351)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        6660..6678
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    6679..6703
                     /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)."
                     /note="color: #31849b"
     RBS             6718..6740
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
ORIGIN
        1 atgggtcatc accatcatca tcacgggtcg gactcagaag tcaatcaaga agctaagcca
       61 gaggtcaagc cagaagtcaa gcctgagact cacatcaatt taaaggtgtc cgatggatct
      121 tcagagatct tcttcaagat caaaaagacc actcctttaa gaaggctgat ggaagcgttc
      181 gctaaaagac agggtaagga aatggactcc ttaagattct tgtacgacgg tattagaatt
      241 caagctgatc aggcccctga agatttggac atggaggata acgatattat tgaggctcac
      301 cgcgaacaga ttggaggtgg agatttgccg attatgcacg atagtgatag gtatgaactc
      361 gtcaaggata ttggctccgg taattttgga gttgcgagat tgatgagaga caagcaaagt
      421 aatgagcttg ttgctgttaa atatatcgag agaggtgaga agatagatga aaatgtaaaa
      481 agggagataa tcaaccacag gtccttaaga catcccaata tcgttagatt caaagaggtt
      541 atattaacac caacccattt agccattgtt atggaatatg catctggagg agaacttttc
      601 gagcgaatct gcaatgcagg ccgcttcagc gaagacgagg cgaggttttt cttccagcaa
      661 ctcatttcag gagttagtta ctgtcatgct atgcaagtat gtcaccgaga cttaaagctc
      721 gagaatacgt tattagatgg tagcccggcc cctcgtctaa agatatgtga tttcggatat
      781 tctaagtcat cagtgttaca ttcgcaacca aaatcaactg ttggaactcc tgcttacatc
      841 gctcctgagg ttttactaaa gaaagaatat gatggaaagg ttgcagatgt ttggtcttgt
      901 ggggttactc tgtatgtcat gctggttgga gcatatcctt tcgaagatcc cgaggaacca
      961 aagaatttca ggaaaactat acatagaatc ctgaatgttc agtatgctat tccggattat
     1021 gttcacatat ctcctgaatg tcgccatttg atctccagaa tatttgttgc tgaccctgca
     1081 aagaggatat caattcctga aataaggaac catgaatggt ttctaaagaa tctaccggca
     1141 gatctaatga acgataacac gatgaccact cagtttgatg aatcggatca accgggccaa
     1201 agcatagaag aaattatgca gatcattgca gaagcaactg ttcctcctgc aggcactcag
     1261 aatctgaacc attacctcac aggaagcttg gacatagatg acgatatgga ggaagactta
     1321 gagagcgacc ttgatgatct tgacatcgac agtagcggag agattgtgta cgcaatgtga
     1381 gtcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     1441 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1501 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     1561 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1621 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1681 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1741 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1801 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1861 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1921 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1981 attttaacaa aatattaacg tttacaattt caggtggcac ttttcgggga aatgtgcgcg
     2041 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2101 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2161 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2221 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2281 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2341 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2401 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2461 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2521 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2581 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2641 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     2701 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2761 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2821 ggtttattgc tgataaatct ggagccggtg agcgtggttc tcgcggtatc attgcagcac
     2881 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2941 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3001 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3061 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3121 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3181 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3241 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3301 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3361 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3421 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3481 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3541 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3601 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3661 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3721 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3781 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3841 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3901 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3961 ttctccttac gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat
     4021 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     4081 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4141 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4201 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     4261 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4321 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4381 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4441 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4501 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4561 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4621 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4681 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4741 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     4801 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     4861 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     4921 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     4981 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     5041 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     5101 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     5161 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5221 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5281 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5341 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5401 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     5461 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5521 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5581 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5641 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     5701 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     5761 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     5821 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     5881 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     5941 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     6001 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     6061 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     6121 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     6181 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     6241 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6301 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6361 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     6421 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6481 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6541 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     6601 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     6661 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     6721 gtttaacttt aagaaggaga tatacc
//
