LOCUS       40924_41420        5597 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSUMO3_ck5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5597)
  AUTHORS   Kegler C.
  TITLE     A multifunctional enzyme is involved in bacterial ether lipid 
            biosynthesis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5597)
  AUTHORS   Kegler C.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAR-2014) Molekulare Biotechnologie / Institut fur 
            Molekulare Biowissenschaften, Goethe Universtaet Frankfurt, 
            Max-von-Laue Strasse 9, Frankfurt am Main, Hessen 60438, Germany
REFERENCE   3  (bases 1 to 5597)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5597)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-MAR-2014) Molekulare Biotechnologie / Institut fur Molekulare 
            Biowissenschaften, Goethe Universtaet Frankfurt, Max-von-Laue 
            Strasse 9, Frankfurt am Main, Hessen 60438, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: ??? v. ???
            Sequencing Technology :: unknown
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..24
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             31..306
                     /codon_start=1
                     /label=SUMO3
                     /note="human small ubiquitin-related modifier 3"
                     /translation="LQEEKPKEGVKTENDHINLKVAGQDGSVVQFKIKRHTPLSKLMKA
                     YCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGG"
     CDS             371..388
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      455..502
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      539..994
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1090..1902)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2024..2612
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2798..2940)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3045..3233)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(4008..4029)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4045..5124)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5125..5202)
                     /label=lacI promoter
     promoter        5511..5529
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5530..5554
                     /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)."
     RBS             5569..5591
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atgggtcatc accatcatca tcacgggtcc ctgcaggagg agaagcccaa ggagggtgtg
       61 aagacagaga atgaccacat caacctgaag gtggccgggc aggacggctc cgtggtgcag
      121 ttcaagatca agaggcacac gccgctgagc aagctgatga aggcctactg cgagaggcag
      181 ggcttgtcaa tgaggcagat cagattcagg ttcgacgggc agccaatcaa tgaaactgac
      241 actccagcac agctggagat ggaggacgag gacaccatcg acgtgttcca gcagcagacg
      301 ggaggtagta ctggatccca cgtgactagt ctcgagggta ccaagcttga attccaattg
      361 actaggtgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
      421 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      481 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
      541 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      601 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      661 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      721 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      781 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      841 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      901 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      961 gcgaatttta acaaaatatt aacgtttaca atttcaggtg gcacttttcg gggaaatgtg
     1021 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     1081 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     1141 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     1201 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     1261 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     1321 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     1381 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     1441 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     1501 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     1561 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     1621 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     1681 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     1741 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     1801 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     1861 tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     1921 gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt
     1981 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2041 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2101 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2161 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2221 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2281 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2341 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2401 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2461 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2521 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2581 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2641 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2701 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2761 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     2821 cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga
     2881 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     2941 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3001 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3061 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     3121 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     3181 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     3241 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     3301 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     3361 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     3421 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     3481 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     3541 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     3601 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     3661 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     3721 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     3781 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     3841 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     3901 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     3961 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     4021 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4081 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     4141 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     4201 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     4261 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     4321 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     4381 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     4441 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     4501 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     4561 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     4621 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     4681 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     4741 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     4801 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     4861 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     4921 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     4981 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     5041 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     5101 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     5161 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     5221 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     5281 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     5341 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     5401 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     5461 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
     5521 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     5581 taagaaggag atatacc
//
