LOCUS       Exported                7565 bp ds-DNA     circular SYN 21-FEB-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7565)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Mar 8, 2019 from SnapGene Viewer 4.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7565
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    104..962
                     /label=HO downstream sequence
                     /note="HO downstream sequence"
     gene            995..1404
                     /label=TEFmut7
     CDS             1405..2535
                     /codon_start=1
                     /gene="cggR"
                     /product="cggR TR"
                     /label=cggR
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDHPFTMNQLIQAQK
                     KLLPDLLLVMQKRFEILQYIRLTEPIGRRSLSASLGISERVLRGEVQFLKEQNLVDIKT
                     NGMTLTEEGYELLSVLEDTMKDVLGLTLLEKTLKERLNLKDAIIVSGDSDQSPWVKKEM
                     GRAAVACMKKRFSGKNIVAVTGGTTIEAVAEMMTPDSKNRELLFVPARGGLGEDVKNQA
                     NTICAHMAEKASGTYRLLFVPGQLSQGAYSSIIEEPSVKEVLNTIKSASMLVHGIGEAK
                     TMAQRRNTPLEDLKKIDDNDAVTEAFGYYFNADGEVVHKVHSVGMQLDDIDAIPDIIAV
                     AGGSSKAEAIEAYFKKPRNTVLVTDEGAAKKLLRDE"
     misc_feature    1405..1512
                     /label=N-terminal tagging sequence
     misc_feature    1417..1434
                     /label=6xHis tag
     misc_feature    1474..1497
                     /label=Xpress Epitope
     misc_feature    1483..1497
                     /label=EK recognition site
     misc_feature    1513..2535
                     /label=codon optimized cggR
     CDS             complement(2547..2758)
                     /gene="lacZ''"
                     /product="5'-beta-galactosidase alpha fragment"
                     /label=lacZ''
                     /note="lacZ''"
     misc_feature    2802..2835
                     /label=loxP site
                     /note="loxP site"
     CDS             3234..3614
                     /codon_start=1
                     /transl_table=11
                     /gene="ble"
                     /product="bleomycin resistance protein"
                     /label=ble
                     /note="ble"
                     /db_xref="GI:11344913"
                     /protein_id="AAG34545.1"
                     /translation="MADQATPNLPSRDFDSTAAFYERLGFGIVFRDAGWMILQRGDLKL
                     EFFAHPGLDPLASWFSCCLRLDDLAEFYRQCKSVGIQETSSGYPRIHAPELQEWGGTMA
                     ALVDPDGTLLRLIQNELLAGIS"
     misc_feature    3880..3913
                     /label=loxP site
                     /note="loxP site"
     misc_feature    4001..4474
                     /label=HO upstream sequence
                     /note="HO upstream sequence"
     primer_bind     complement(4465..4482)
                     /label=SEQ_4_REV
     misc_feature    6502..7184
                     /label=colE1
                     /note="colE1"
ORIGIN
        1 atgaccatga ttacgccact agtccgaggc ctcgagatcc gatatcgccg tggcggccgc
       61 cagctgaagc ttaattatcc tgggcacgag tgaaacaaag ctaaaacctt tatttagcat
      121 ggccattgaa tgtaacaatt atatatatcg caagcacaaa aaatcaagga gagagaacta
      181 ccactttgtt catgtgtaca atgttcatta tctccataag caaaaaaaaa aaatagaaaa
      241 catatgctat aaggttgata ttctcacgag taagcggcac ttgctactta ttgacattgc
      301 agatttttgg ctacagaaat agtatattag agattataat tgctaatcaa atcaaaatat
      361 aaaattagta aaccaaacca tttataccct tccttagtag ttatggattg ttttttaatg
      421 atatttctgc aaaccaaaga aagattgtta tccagataga atttagtttt gatattcatt
      481 tttttgttga agattgaacg ccatatctgg gcctcataat tcaaaagacg gtgccattat
      541 cggtagcgtt tcgcattgta ctggatttca gaaatttcac agttgatgaa tcgaaaagaa
      601 tggtctcatt gcaacacgta aggttaagat gtcccttttt accattatag gcaataaatg
      661 aatcataaaa cgaccgtata ctggtgaaat agtagggaga acgagtacct gtagtaaaaa
      721 gtataaatca tagttaatcg ggcaatgtcc ctcgatcaag gagtattgtg tcatgttcga
      781 gacaaacgcc aacatttttg tttcttttgg acaaatgttg tttgcattta tgatccgtta
      841 tattttgatc taatgtagag ttgcacgtag ttcttactgg caaagaaatc gatgcatacc
      901 aaaaaagaat aaaggtgata tttgatcttt accgtttagt tccaacgtaa aattgtgcct
      961 ttggacttaa aatggcgtcg tacgctaggt cgagatagct tcaaaatgtc tctactcctt
     1021 ttttactctt ccagattttc tcggactccg cgcaccgccg taccacttca aaacacccaa
     1081 gcacagcata ctaaattccc cctcctcctt cctctagggt gccgttaatt acccgtacta
     1141 aaggtttgga aaaggaaaaa gagaccgcct cgtccctttt tcttcgtcgg agaaggcaat
     1201 aaaaattttt atcacgtttc tttctcttga aaactttttt ttcgattttg ttctctttcg
     1261 acgacctccc attgatattt gagttaacaa acggtcttca atttctcaag tttcagcttc
     1321 atttttcctg ttctattaca acttttttta cttcttgctc attggaaaga aagcatagca
     1381 atctaatcta agttttctag aaaaatgcgg ggttctcatc atcatcatca tcatggtatg
     1441 gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
     1501 catcccttca ccatgaatca attgatccaa gcccaaaaga agttgttgcc agatttgttg
     1561 ttggtcatgc aaaagagatt cgaaatcttg caatatatca gattgaccga acctatcggt
     1621 agaagatctt tgtctgcttc tttgggtatc tccgaaagag ttttgagagg tgaagtccaa
     1681 ttcttgaaag aacaaaactt ggtcgacatc aagactaacg gtatgacttt gactgaagaa
     1741 ggttacgaat tattgtccgt cttggaagat accatgaagg atgttttggg tttgaccttg
     1801 ttggaaaaga ctttgaaaga aagattgaac ttgaaggacg ccatcatcgt ttctggtgat
     1861 tctgatcaat ctccatgggt caaaaaagaa atgggtagag ctgctgttgc ttgcatgaag
     1921 aaaagatttt ctggtaagaa catcgttgct gttactggtg gtactactat tgaagctgtt
     1981 gctgaaatga tgaccccaga ttctaagaac agagaattgt tgtttgttcc agctagaggt
     2041 ggtttgggtg aagatgttaa gaatcaagct aacaccattt gcgctcatat ggctgaaaaa
     2101 gcttcaggta cttacagatt gttattcgtc ccaggtcaat tgtctcaagg tgcttactct
     2161 tccattatcg aagaaccatc tgtcaaagaa gtcttgaaca ccatcaaatc cgcttccatg
     2221 ttggttcacg gtattggtga agctaaaact atggctcaaa gaagaaacac cccattggaa
     2281 gatttgaaaa agatcgatga taacgatgct gttaccgaag ctttcggtta ctactttaat
     2341 gctgatggtg aagttgttca caaggttcat tcagttggta tgcaattgga tgatattgat
     2401 gccatcccag atattattgc agttgctggt ggttcttcta aggctgaagc aattgaagct
     2461 tacttcaaga agccaagaaa cactgttttg gttactgatg aaggtgctgc caaaaagttg
     2521 ttgagagatg aatgatagct cgaggtaccg agctcgaatt cactggccgt cgttttacaa
     2581 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2641 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2701 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtacg
     2761 ccagctgaag cttcgtacgc tgcaggtcga caacccttaa tataacttcg tataatgtat
     2821 gctatacgaa gttattaggt ctagagatct gtttagcttg cctcgtcccc gccgggtcac
     2881 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
     2941 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
     3001 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
     3061 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
     3121 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
     3181 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatggccg
     3241 accaagcgac gcccaacctg ccatcacgag atttcgattc cacggccgcc ttctatgaaa
     3301 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3361 tcaagctgga gttcttcgcc caccccgggc tcgatcccct cgcgagttgg ttcagctgct
     3421 gcctgaggct ggacgacctc gcggagttct accggcagtg caaatccgtc ggcatccagg
     3481 aaaccagcag cggctatccg cgcatccatg cccccgaact gcaggagtgg ggaggcacga
     3541 tggccgcttt ggtcgacccg gacgggacgc tcctgcgcct gatacagaac gaattgcttg
     3601 caggcatctc atgatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     3661 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     3721 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     3781 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     3841 ccatccagtg tcgaaaacga gctctcgaga acccttaata taacttcgta taatgtatgc
     3901 tatacgaagt tattaggtga tatcagatcc actagtggcc tatgcgcgcc gccatccagt
     3961 gtcgaaaacg agctcgaatt cctgggggaa caacttcaca gaatgttttg tcatattgtc
     4021 gaagtggtca caaaacaaga gaagttccgc caattataaa aagggaaccc gtatatttca
     4081 gcttcacgga tgatttccag ggtgagagta ctgtatatgg gcttacgata gaaggccata
     4141 aaaatttctt gcttggcaac aaaatagaag tgaaatcatg tcgaggctgc tgtgtgggag
     4201 aacagcttaa aatatcacaa aaaaagaatc taaaacactg tgttgcttgt cccagaaagg
     4261 gaatcaagta tttttataaa gattggagtg gtaaaaatcg agtatgtgct agatgctatg
     4321 gaagatacaa attcagcggt catcactgta taaattgcaa gtatgtacca gaagcacgtg
     4381 aagtgaaaaa ggcaaaagac aaaggcgaaa aattgggcat tacgcccgaa ggtttgccag
     4441 ttaaaggacc agagtgtata aaatgtggcg gaatcttaca gtggcctatg cggccgctct
     4501 agaactagtg gatcgatccc caattcgccc tatagtgagt cgtattacaa ttcactggcc
     4561 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4621 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4681 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     4741 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4801 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4861 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4921 gtcaagctct aaatcggggt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4981 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5041 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5101 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5161 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5221 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5281 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5341 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5401 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5461 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5521 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5581 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5641 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5701 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5761 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5821 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5881 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5941 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6001 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6061 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6121 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6181 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6241 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6301 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6361 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6421 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6481 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6541 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6601 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6661 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6721 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6781 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6841 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6901 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6961 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7021 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7081 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7141 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7201 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7261 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7321 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     7381 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7441 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     7501 tgcttccgcg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     7561 cagct
//
