LOCUS       40924_41415        5585 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSUMO3_ck4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5585)
  AUTHORS   Kegler C.
  TITLE     A multifunctional enzyme is involved in bacterial ether lipid 
            biosynthesis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5585)
  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 5585)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5585)
  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            "Unpublished"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..5585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        164..182
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    183..207
                     /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             222..244
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             257..274
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             281..556
                     /codon_start=1
                     /label=SUMO3
                     /note="human small ubiquitin-related modifier 3"
                     /translation="MSEEKPKEGVKTENDHINLKVAGQDGSVVQFKIKRHTPLSKLMKA
                     YCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGG"
     CDS             609..626
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      693..740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      777..1232
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1328..2140)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2262..2850
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3036..3178)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3283..3471)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(4246..4267)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4283..5362)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5363..5440)
                     /label=lacI promoter
ORIGIN
        1 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
       61 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
      121 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
      181 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
      241 gagatatacc atgggtcatc accatcatca tcacggaggt atgtccgagg agaagcccaa
      301 ggagggtgtg aagacagaga atgaccacat caacctgaag gtggccgggc aggacggctc
      361 cgtggtgcag ttcaagatca agaggcacac gccgctgagc aagctgatga aggcctactg
      421 cgagaggcag ggcttgtcaa tgaggcagat cagattcagg ttcgacgggc agccaatcaa
      481 tgaaactgac actccagcac agctggagat ggaggacgag gacaccatcg acgtgttcca
      541 gcagcagacg ggaggtagta ctggatccca cgtgactagt ctcgagggta cccaattgac
      601 taggtgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
      661 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      721 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
      781 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      841 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      901 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      961 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1021 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1081 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1141 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1201 gaattttaac aaaatattaa cgtttacaat ttcaggtggc acttttcggg gaaatgtgcg
     1261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     1321 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     1381 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1441 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1501 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1561 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     1621 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     1681 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     1741 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     1801 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     1861 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     1921 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     1981 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     2041 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     2101 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2161 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     2221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2401 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2641 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2941 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3001 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     3061 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     3121 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3181 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3241 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3301 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     3361 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     3421 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     3481 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     3541 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     3601 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     3661 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     3721 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     3781 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     3841 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     3901 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     3961 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4021 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4081 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     4141 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     4201 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     4261 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4321 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     4381 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     4441 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     4501 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     4561 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     4621 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     4681 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     4741 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     4801 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     4861 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     4921 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     4981 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     5041 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     5101 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     5161 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     5221 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     5281 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     5341 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     5401 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     5461 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     5521 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     5581 gccac
//
