LOCUS       62056_22370        7395 bp DNA     circular SYN 28-JAN-2020
DEFINITION  Cloning vector pUD1074, complete sequence.
ACCESSION   MN498128
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7395)
  AUTHORS   Valk LC.
  TITLE     'Candidatus Galacturonibacter soehngenii' shows hetero-acetogenic 
            catabolism of galacturonic acid but lacks a canonical carbon 
            monoxide dehydrogenase/acetyl-CoA synthase complex
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7395)
  AUTHORS   Valk LC.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-SEP-2019) Biotechnology, Delft University of 
            Technology, Van der Maasweg 9, Delft, Zuid Holland 2629 HZ, 
            Netherlands
REFERENCE   3  (bases 1 to 7395)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-SEP-2019) Biotechnology, Delft University of Technology, Van der
            Maasweg 9, Delft, Zuid Holland 2629 HZ, Netherlands"
FEATURES             Location/Qualifiers
     source          1..7395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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             267..289
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             300..317
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             318..359
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             360..380
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             396..2033
                     /codon_start=1
                     /transl_table=11
                     /product="hybrid cluster protein"
                     /label=hybrid cluster protein
                     /note="hydroxylamine reductase; putative novel carbon
                     monoxide dehydrogenase; codon optimized for Escherichia 
                     coli"
                     /protein_id="QHN70777.1"
                     /translation="MENKMFCYQCQETAGCTGCTQMGVCGKTPEVAGMQDLLVYVTKGL
                     AEVTTKLRKENVAIEKSTNHYITWNLFTTITNANFDEAVFVERINKSLDIKKELLAKLS
                     DKSGLSDAAIWEDYSVEGMRAKAAQVGVLATENEDVRSLRELITYGLKGLCAYMKHANA
                     LSKENEAVDIFVQETLAKLLDDSLSVNELIALTLETGKYGVDGMALLDEANTSSYGNPE
                     ITSVNIGVGKNPGILISGHDLRDMEMLLEQTKGTGVDVYTHSEMLPAHYYPAFKKYDHF
                     VGNYGNAWWKQKEEFESFNGPILMTTNCIVPPKASYKDRLFTTGSAGMEGCKHIEGNEG
                     EVKDFSEIIALAKTCQPPTEIETGSIVGGFAHNQVFALADKVVDAVKSGAIKKFVVMAG
                     CDGRAKSRNYYTDFAKALPNDSVILTAGCAKYKYNKLNLGDIGGIPRVLDAGQCNDSYS
                     LVLIALKLQEVFGLESVNDLPIAYNIAWYEQKAVIVLLSLLHLGVKNIHLGPTLPAFLS
                     PNVVNVLVENFGIAGITDVENDLKILLA"
     terminator      2110..2157
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2481..2585
                     /label=AmpR promoter
     CDS             2586..3443
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3617..4205
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4391..4530)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4635..4823)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(6101..6122)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6138..7217)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7218..7295)
                     /label=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 tcaccatgga aaacaagatg ttctgctacc
      421 agtgtcaaga aaccgcaggt tgtaccggtt gcacccagat gggtgtttgt ggtaaaacac
      481 cggaagttgc aggtatgcag gatctgctgg tttatgttac caaaggtctg gcagaagtta
      541 ccaccaaact gcgtaaagaa aatgtggcca ttgaaaaaag caccaaccat tatatcacct
      601 ggaacctgtt taccaccatt accaatgcca attttgatga agccgttttt gtggaacgca
      661 tcaataaaag cctggacatc aaaaaagaac tgctggcaaa actgagcgat aaaagcggtc
      721 tgagtgatgc agcaatttgg gaagattata gcgttgaagg tatgcgtgca aaagcagcac
      781 aggttggtgt tctggcaacc gaaaatgaag atgttcgtag cctgcgtgaa ctgattacct
      841 atggtctgaa aggtctgtgt gcctatatga aacatgcaaa tgcactgagc aaagaaaacg
      901 aagccgttga tatttttgtg caagaaaccc tggccaaact gctggatgat agcctgagcg
      961 ttaacgaact gattgcactg accctggaaa ccggtaaata tggtgttgat ggtatggcac
     1021 tgctggacga agcaaatacc agcagctatg gtaatccgga aattaccagc gttaatattg
     1081 gtgtgggtaa aaatccgggt attctgatta gcggtcatga tctgcgtgat atggaaatgc
     1141 tgctggaaca gaccaaaggc accggtgtgg atgtttatac ccatagcgaa atgttaccgg
     1201 cacattatta tccggcattc aaaaagtatg atcacttcgt gggtaattat ggtaatgcct
     1261 ggtggaaaca gaaagaagaa tttgaaagct ttaacggtcc gattctgatg accaccaatt
     1321 gtattgttcc gcctaaagcc agctataaag atcgtctgtt tacaaccggt agcgcaggta
     1381 tggaaggttg caaacacatt gaaggtaatg aaggcgaagt gaaagatttt agcgaaatca
     1441 ttgcactggc caaaacctgt cagcctccga ccgaaattga aaccggcagc attgttggtg
     1501 gttttgcaca taatcaggtt tttgccctgg cagataaagt tgttgatgca gtgaaaagcg
     1561 gtgccatcaa aaagtttgtt gttatggcag gttgtgatgg tcgtgcaaaa agccgtaact
     1621 attataccga ttttgccaaa gcactgccga atgatagcgt tattctgacc gcaggctgtg
     1681 caaaatacaa gtataacaaa ctgaacctgg gtgacattgg tggtattccg cgtgttctgg
     1741 atgcaggtca gtgtaatgat agctatagcc tggttctgat tgccctgaaa ctgcaagaag
     1801 tttttggtct ggaaagcgtt aatgatctgc cgattgcata taacattgcc tggtatgaac
     1861 agaaagccgt tattgttctg ctgagcctgc tgcatctggg cgttaaaaac attcatctgg
     1921 gtccgacact gcctgccttt ctgagcccga atgttgttaa tgttctggtt gaaaattttg
     1981 gcattgccgg tattaccgat gtggaaaacg atctgaaaat tctgctggcc taaaagggcg
     2041 agctcagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2101 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2161 aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc ataaccaagc
     2221 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
     2281 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     2341 agcttatcga tgataagctg tcaaacatga gaattaattc ttgaagacga aagggcctcg
     2401 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2461 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2521 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2581 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2641 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2701 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2761 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2821 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2881 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2941 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3001 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3061 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3121 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3181 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3241 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3301 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3361 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3421 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3481 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3541 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3601 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3661 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3721 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3781 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3841 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3901 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3961 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4021 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4081 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4141 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4201 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4261 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4321 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4381 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4441 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     4501 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4561 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4621 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     4681 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4741 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4801 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4861 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4921 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4981 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     5041 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     5101 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     5161 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     5221 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     5281 cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg
     5341 gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag
     5401 aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca
     5461 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
     5521 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
     5581 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
     5641 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
     5701 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
     5761 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
     5821 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     5881 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     5941 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     6001 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     6061 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     6121 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6181 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     6241 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     6301 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     6361 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     6421 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     6481 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     6541 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     6601 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     6661 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     6721 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     6781 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     6841 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     6901 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     6961 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     7021 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     7081 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     7141 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     7201 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     7261 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     7321 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     7381 aaggaatggt gcatg
//
