LOCUS       Exported                4002 bp ds-DNA     circular SYN 15-NOV-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4002)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 3, 2020 from SnapGene 4.3.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4002
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     41..71
                     /label=oWL746
                     /note="Description Gene Use Legacy Name"
     primer_bind     41..60
                     /label=T7
     primer_bind     41..60
                     /label=oWL228
                     /note="Description:T7_f Gene:T7 promotor Use:Sequencing 
                     Legacy Name:T7_f"
     promoter        41..59
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(89..129)
                     /label=oWL721
                     /note="Description Gene Use Legacy Name"
     RBS             97..119
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             127..158
                     /label=His-N-DHFR
     CDS             133..150
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     133..150
                     /label=oWL826
                     /note="Description:His Tag forward Gene Use:Sequencing 
                     Legacy Name"
     CDS             160..1260
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     160..184
                     /label=oWL836
                     /note="Description:C-MBP-Fw Gene:MBP Use:Cloning Legacy 
                     Name"
     primer_bind     160..184
                     /label=oWL677
                     /note="Description:Fw_C-MBP Gene:MBP Use:Cloning Legacy 
                     Name"
     primer_bind     198..221
                     /label=oWL334
                     /note="Description Gene Use Legacy Name"
     primer_bind     complement(305..324)
                     /label=oWL335
                     /note="Description Gene Use Legacy Name"
     primer_bind     681..702
                     /label=oWL448
                     /note="Description:MBP_for. Gene:MBP Use:Sequencing Legacy 
                     Name"
     primer_bind     1234..1256
                     /label=oWL133
                     /note="MBP_F"
     primer_bind     complement(1240..1260)
                     /label=oWL835
                     /note="Description:C-MBP-Rv Gene:MBP Use:Cloning Legacy 
                     Name"
     primer_bind     1266..1294
                     /label=oWL623
                     /note="Description:SNAP_no_Bam_f Gene Use Legacy Name"
     primer_bind     1268..1292
                     /label=oWL740
                     /note="Description Gene Use Legacy Name"
     primer_bind     1269..1287
                     /label=oWL736
                     /note="Description Gene Use Legacy Name"
     CDS             1270..1293
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     1270..1292
                     /label=oWL744
                     /note="Description Gene Use Legacy Name"
     primer_bind     complement(1272..1293)
                     /label=oWL678
                     /note="Description:Rv_C-MBP Gene:MBP Use:Cloning Legacy 
                     Name"
     primer_bind     complement(1272..1293)
                     /label=oWL013
                     /note="Spo11_GST_rev"
     primer_bind     complement(1272..1293)
                     /label=oWL016
                     /note="Spo11_38_GST_rev"
     misc_feature    1279..1299
                     /label=fwd primer overhang
     primer_bind     complement(1330..1354)
                     /label=oWL640
                     /note="AB5"
     misc_feature    1355..1379
                     /label=rev primer overhang
     primer_bind     1361..1390
                     /label=oWL734
                     /note="Description Gene Use Legacy Name"
     primer_bind     1361..1380
                     /label=oWL772
                     /note="Description:CTag_backbone_fwd Gene Use:Cloning 
                     Legacy Name"
     primer_bind     complement(1448..1466)
                     /label=oWL227
                     /note="Description:STO720 (T7r) Gene:T7 terminator (pST44) 
                     Use:Sequencing Legacy Name:STO720"
     terminator      1462..1509
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1473..1515)
                     /label=oWL745
                     /note="Description Gene Use Legacy Name"
     promoter        1574..1665
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2164..2192
                     /label=oWL481
                     /note="Description:Amp_fwd Gene Use:Mutagenesis Legacy 
                     Name"
     primer_bind     complement(2164..2192)
                     /label=oWL480
                     /note="Description:Amp_rev Gene Use:Mutagenesis Legacy 
                     Name"
     terminator      2530..2569
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2600..3188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3374..3513
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3615..3806)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cagatgtagg tgttggcacc atgcatcgat cccgcgaaat taatacgact cactataggg
       61 agaccacaac ggtttccctc cggctccgga aataattttg tttaacttta agaaggagat
      121 atacatatgg cacaccatca ccaccatcac agcagcggca tgaaaatcga agaaggtaaa
      181 ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa
      241 ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa
      301 ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc
      361 tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag
      421 gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac
      481 ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca
      541 aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg
      601 ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt
      661 tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct
      721 ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca
      781 gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc
      841 aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta
      901 ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt
      961 aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact
     1021 gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct
     1081 tacgaggaag agttggcgaa agatccacgt attgccgcca ctatggaaaa cgcccagaaa
     1141 ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg
     1201 gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact
     1261 agcagcggtc tggaagttct gttccagggg cccggatccg tcgacgagct cactagtcgc
     1321 ggccgctttc gaatctagag cctgcagtct cgacaagctt gtcgagaagt actagaggat
     1381 gtacaacgcg tgcgcgctga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1441 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1501 ggttttttgc tgaaaggagg aactatatcg ggattagccc gcctaatgag cgggcttttt
     1561 tttaattccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1621 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1681 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1741 gaaacgctcg tgaaagtaaa agacgcagag gaccaattgg gggcacgagt gggatacata
     1801 gaactggact tgaatagcgg taaaatcctt gagagttttc gccctgaaga gcgttttcca
     1861 atgatgagca ctttcaaagt tctgctatgt ggagcagtat tatcccgtgt agatgcgggg
     1921 caagagcaac tcggacgacg aatacactat tcgcagaatg acttggttga atactcccca
     1981 gtgacagaaa agcaccttac ggacggaatg acggtaagag aattatgtag tgccgccata
     2041 acgatgagtg ataacactgc ggcgaactta cttctgacaa ccatcggtgg accgaaggaa
     2101 ttaaccgctt ttttgcacaa tatgggagac catgtaactc gccttgaccg ttgggaacca
     2161 gaactgaatg aagccatacc aaacgacgag cgagacacca caatgcctgc ggcaatggca
     2221 acaacattac gcaaactatt aactggcgaa ctacttactc tggcttcacg gcaacaatta
     2281 atagactggc ttgaagcgga taaagttgca ggaccactac tgcgttcggc acttcctgct
     2341 ggctggttta ttgctgataa atctggggca ggagagcgtg gttcacgggg tatcattgcc
     2401 gcacttggac cagatggtaa gccttcccgt atcgtagtta tctacacgac gggtagtcag
     2461 gcaactatgg acgaacgaaa tagacagatt gctgaaatag gggcttcact gattaagcat
     2521 tggtaaaccg atacaattaa aggctccttt tggagccttt ttttttggac ggaccggtag
     2581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2701 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     2761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3241 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3301 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3361 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3421 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3481 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     3541 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3601 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     3661 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     3721 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     3781 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     3841 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     3901 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     3961 agagaaaaat cactcagggt caatgccagc gcttcgttaa ta
//
