LOCUS       V009586                 6476 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009586
VERSION     V009586
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   Keys TG, Wetter M, Hang I, Rutschmann C, Russo S, Mally M, Steffen
            M, Zuppiger M, Muller F, Schneider J, Faridmoayer A, Lin CW, Aebi M.
  TITLE     A biosynthetic route for polysialylating proteins in Escherichia
            coli
  JOURNAL   Metab. Eng. (2017) In press
   PUBMED   29101090
REFERENCE   2  (bases 1 to 6476)
  AUTHORS   Keys TG.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-NOV-2016) Institute of Microbiology, ETH Zurich,
            Vladimir-Prelog-Weg 4, Zurich, ZH 8093, Switzerland
REFERENCE   3  (bases 1 to 6476)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6476)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            (2017) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (21-NOV-2016) Institute of Microbiology, ETH Zurich,
            Vladimir-Prelog-Weg 4, Zurich, ZH 8093, Switzerland"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      28..75
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(298..954)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(955..1057)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(1583..2127)
                     /direction=LEFT
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     protein_bind    complement(2318..2339)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2355..3434)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(3435..3512)
                     /label="lacI promoter"
     promoter        3550..3580
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    3584..3608
                     /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)."
     CDS             3656..4486
                     /codon_start=1
                     /product="LgtB"
                     /label="LgtB"
                     /protein_id="ATN32183.1"
                     /translation="MSQNHVISLASAAERRAHIADTFGRHGIPFQFFDALMPSERLEQA
                     MAELVPGLSAHPYLSGVEKACFMSHAVLWKQALDEGLPYITVFEDDVLLGEGAEKFLAE
                     DAWLQERFDPDTAFIVRLETMFMHVLTSPSGVADYCGRAFPLLESEHWGTAGYIISRKA
                     MRFFLDRFAALPPEGLHPVDLMMFSDFFDREGMPVCQLNPALCAQELHYAKFHDQNSAL
                     GSLIEHDRLLNRKQQRRDSPANTFKHRLIRALTKISREREKRRQRREQFIVPFQ"
     promoter        4498..4528
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an ""up"" mutation"
     regulatory      4498..4503
                     /regulatory_class="minus_35_signal"
     regulatory      4522..4528
                     /note="-10 seq (TATAAT; Pribnow box)"
                     /regulatory_class="minus_10_signal"
     protein_bind    4532..4556
                     /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)."
     misc_feature    4534..4554
                     /label="lac operator"
                     /note="lac operator"
     CDS             4604..6463
                     /note="UDP-glucose:protein N-beta-glucosyltransferase from
                     Actinobacillus pleuropneumoniae serotype 7 (strain AP76).
                     Accession#: B3H2N2"
                     /label="UDP-glucose:protein N-beta-glucosyltransferase"
ORIGIN
        1 taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
       61 ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg
      121 gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct
      181 gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac
      241 ttattcaggc gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc
      301 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
      361 gccatcacag acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
      421 cgtataatat ttgcccatag tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
      481 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
      541 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
      601 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
      661 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
      721 tttcattgcc atacggaact ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
      781 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
      841 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
      901 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
      961 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     1021 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt
     1081 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
     1141 ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat
     1201 ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct
     1261 cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct
     1321 agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca
     1381 tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat
     1441 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
     1501 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     1561 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
     1621 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1681 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     1741 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     1801 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     1861 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     1921 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     1981 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     2041 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     2101 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     2161 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     2221 aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     2281 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     2341 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2401 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     2461 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     2521 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     2581 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca
     2641 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     2701 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     2761 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     2821 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     2881 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     2941 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     3001 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     3061 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     3121 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     3181 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     3241 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     3301 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     3361 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     3421 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     3481 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttacataa
     3541 acgctagtct ttacacttta tgcttccggc tcgtataatg tgtggaattg tgagcggata
     3601 acaattcccc actagaaata actctttaat actatttaca taaggaggcg agaacatgtc
     3661 tcaaaaccat gtgatctctt tggcatctgc ggccgagcgt cgcgcgcaca tcgcggatac
     3721 cttcggccgt catggcattc catttcaatt ttttgacgcg ctcatgccat ccgaacgttt
     3781 agagcaagcg atggcggaac tggtgccagg tttgtccgcg catccgtacc tcagcggcgt
     3841 tgagaaagct tgttttatgt cgcacgccgt tctgtggaag caggcgttag acgagggtct
     3901 gccgtatatt accgttttcg aggacgacgt tttgttgggt gaaggtgcgg aaaagttttt
     3961 ggctgaggac gcttggctgc aggagcgttt tgatcctgat acggccttca ttgttcgcct
     4021 ggaaaccatg ttcatgcatg tcttgacctc cccgagcggt gtcgccgact actgtggtcg
     4081 cgctttcccg ttgctggaga gcgagcactg gggcaccgcg ggttatatta tctcccgtaa
     4141 agcaatgcgt ttcttcctgg atcgcttcgc tgcgctgccg ccggagggct tgcacccggt
     4201 ggacctgatg atgttcagcg attttttcga ccgcgagggt atgcctgttt gccaactgaa
     4261 tccggcgttg tgcgcgcagg agttacacta cgcaaaattc cacgatcaga acagcgcgct
     4321 gggtagcctg atcgagcatg accgtctgct gaaccgcaaa cagcaacgcc gtgacagccc
     4381 ggctaacacc tttaagcacc gtctgatccg cgcgctgact aagatcagcc gtgaacgtga
     4441 gaagcgtcgt cagcgtcgcg aacaattcat tgtgccgttt caataattcg cctaggcttt
     4501 acactttatg cttccggctc gtataatgtg tggaattgtg agcggataac aattccccac
     4561 tagaaataaa cccccagacc cacagatact caggaggtct cgcatggaga acgagaataa
     4621 gcctaatgtt gcaaactttg aggcggcagt tgctgtcaaa gactacgaga aggcttgctc
     4681 cgagctgttg ctgatcttgt ctcagctgga tagcaacttt ggcggtattc aagaaatcga
     4741 gttcgagtac ccggttcaac tccaggacct cgaacaagag aaaattgtat acttctgcac
     4801 ccgcatggcg accgcgatca cgaccttgtt ttccgaccct gttctggaga tcagcgacct
     4861 gggtgttcag cgtttcctgg tttaccaacg ttggctcgca ttgatctttg catcgtcccc
     4921 gtttgttaac gcggatcata ttctgcaaac ctataaccgc gaaccgaacc gtaaaaactc
     4981 tttggaaatc catctggact ctagcaaatc cagcctcatc aaattttgca ttttatactt
     5041 gccggagtcc aacgtcaacc tgaacttgga cgtgatgtgg aacattagcc cggagttgtg
     5101 cgccagcctg tgcttcgcac tgcagtcgcc gcgcttcatc ggcacctcta ccgcgttcaa
     5161 caagcgtgcg actattttac aatggtttcc acgtcacctg gaccagctga aaaatttgaa
     5221 caacattccg agcgcgattt cccacgacgt ctatatgcat tgctcttacg acacctccgt
     5281 gaacaagcat gatgtcaaac gtgcgctgaa ccatgtgatt cgccgtcata tcgagtccga
     5341 atacggctgg aaagaccgtt acgtggcgca catcggctac cgtaacaaca agccggtgat
     5401 ggttgttctg ctggagcact ttcactctgc acactccatc tatcgtaccc actcgacctc
     5461 catgattgcg gcccgcgagc acttttacct gatcggtttg ggtagcccga gcgttgatca
     5521 agcgggccaa gaggtttttg acgagtttca tctcgttgcg ggtgataata tgaagcagaa
     5581 gctggagttc attcgttcgg tgtgcgagtc taacggtgcc gccatcttct acatgccgtc
     5641 cattggcatg gatatgacca ccatcttcgc ttcgaatact cgccttgcgc ctatccaggc
     5701 gatcgcactg ggccaccctg ccaccaccca cagcgacttt atcgagtacg tcatcgttga
     5761 ggacgattac gtgggctccg aggcatgctt tagcgaaacg ttgctgagac tgccgaagga
     5821 cgcgctgccg tatgtcccgt ccgccttggc cccggagaaa gttgattatc ttttgcgtga
     5881 gaatccagag gtcgttaata ttggcattgc gagcaccacc atgaagctga acccttattt
     5941 cttggaagcg ctgaaggcga tccgtgatcg tgctaaagtc aaagttcact tccattttgc
     6001 gctgggccaa agcaacggta ttacccatcc atacgttgag cgtttcatta agtcctacct
     6061 gggtgatagc gcgaccgctc acccacattc cccgtatcac caatacttgc gcatcttgca
     6121 taattgcgac atgatggtta acccgttccc gtttggtaac acgaatggca tcattgatat
     6181 ggtgaccctc ggtctggttg gtgtttgcaa aaccggtgca gaggtccacg aacacatcga
     6241 cgagggcttg tttaagcgtc tgggcttgcc ggaatggctg attgccaaca ccgttgacga
     6301 atatgtggag cgtgcggtgc gtttggcgga aaaccaccag gaacgcttgg agctgcgccg
     6361 ctatattatc gagaataatg gcttgaatac gctgtttacc ggtgacccgc gtccgatggg
     6421 tcaggttttt ctggaaaaat tgaacgcttt cttgaaggag aactaatagc gtctct
//
