LOCUS       Exported                3497 bp ds-DNA     circular SYN 30-JAN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3497)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 3, 2020 from SnapGene 4.3.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3497
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    186..325
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(427..618)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     855..885
                     /label=oWL746
                     /note="Description Gene Use Legacy Name"
     primer_bind     855..874
                     /label=oWL228
                     /note="Description:T7_f Gene:T7 promotor Use:Sequencing 
                     Legacy Name:T7_f"
     promoter        855..873
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(903..943)
                     /label=oWL721
                     /note="Description Gene Use Legacy Name"
     RBS             911..933
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(950..970)
                     /label=oWL738
                     /note="Description Gene Use Legacy Name"
     CDS             953..970
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             989..1534
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     primer_bind     complement(1511..1552)
                     /label=oWL624
                     /note="Description:SNAP_noBAM_rev Gene Use Legacy Name"
     primer_bind     1526..1565
                     /label=oWL623
                     /note="Description:SNAP_no_Bam_f Gene Use Legacy Name"
     CDS             1541..1564
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     1541..1563
                     /label=oWL744
                     /note="Description Gene Use Legacy Name"
     primer_bind     1541..1563
                     /label=oWL740
                     /note="Description Gene Use Legacy Name"
     primer_bind     1541..1558
                     /label=oWL736
                     /note="Description Gene Use Legacy Name"
     primer_bind     1541..1552
                     /label=oWL729
                     /note="Description Gene Use Legacy Name"
     primer_bind     complement(1543..1564)
                     /label=oWL013
                     /note="Spo11_GST_rev"
     primer_bind     complement(1543..1564)
                     /label=oWL016
                     /note="Spo11_38_GST_rev"
     primer_bind     complement(1543..1564)
                     /label=oWL678
                     /note="Description:Rv_C-MBP Gene:MBP Use:Cloning Legacy 
                     Name"
     primer_bind     complement(1587..1611)
                     /label=oWL622
                     /note="Description:pLIB_Strep_mutate_rev Gene Use Legacy 
                     Name"
     primer_bind     complement(1639..1663)
                     /label=oWL640
                     /note="AB5"
     primer_bind     1670..1699
                     /label=oWL734
                     /note="Description Gene Use Legacy Name"
     primer_bind     1670..1689
                     /label=oWL772
                     /note="Description:CTag_backbone_fwd Gene Use:Cloning 
                     Legacy Name"
     primer_bind     complement(1757..1775)
                     /label=oWL227
                     /note="Description:STO720 (T7r) Gene:T7 terminator (pST44) 
                     Use:Sequencing Legacy Name:STO720"
     terminator      1771..1818
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1782..1824)
                     /label=oWL745
                     /note="Description Gene Use Legacy Name"
     promoter        1883..1974
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2473..2501
                     /label=oWL481
                     /note="Description:Amp_fwd Gene Use:Mutagenesis Legacy 
                     Name"
     primer_bind     complement(2473..2501)
                     /label=oWL480
                     /note="Description:Amp_rev Gene Use:Mutagenesis Legacy 
                     Name"
     terminator      2839..2878
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2909..3497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
      241 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      301 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      361 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      421 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
      481 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
      541 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
      601 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tggggtaatg
      661 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
      721 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
      781 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttgg caccatgcat
      841 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctccggctc
      901 cggaaataat tttgtttaac tttaagaagg agatatacat atgggcagca gccatcatca
      961 tcatcatcac agcagcggat ctggatctat ggataaagat tgtgaaatga aacgtaccac
     1021 cctggatagc ccgctgggca aactggaact gagcggctgc gaacagggcc tgcatgaaat
     1081 taaactgctg ggtaaaggca ccagcgcggc cgatgcggtt gaagttccgg ccccggccgc
     1141 cgtgctgggt ggtccggaac cgctgatgca ggcgaccgcg tggctgaacg cgtattttca
     1201 tcagccggaa gcgattgaag aatttccggt tccggcgctg catcatccgg tgtttcagca
     1261 ggagagcttt acccgtcagg tgctgtggaa actgctgaaa gtggttaaat ttggcgaagt
     1321 gattagctat cagcagctgg cggccctggc gggtaatccg gcggccaccg ccgccgttaa
     1381 aaccgcgctg agcggtaacc cggtgccgat tctgattccg tgccatcgtg tggttagctc
     1441 tagcggtgcg gttggcggtt atgaaggtgg tctggcggtg aaagagtggc tgctggccca
     1501 tgaaggtcat cgtctgggta aaccgggtct gggttccgga ctggaagttc tgttccaggg
     1561 gcccggatcc cggtccgaca tatgagcgcg cggaattcaa aggcctacgt cgacgagctc
     1621 actagtcgcg gccgctttcg aatctagagc ctgcagtctc gacaagcttg tcgagaagta
     1681 ctagaggatg tacaacgcgt gcgcgctgat ccggctgcta acaaagcccg aaaggaagct
     1741 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1801 gtcttgaggg gttttttgct gaaaggagga actatatcgg gattagcccg cctaatgagc
     1861 gggctttttt ttaattcccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1921 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1981 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2041 gctcacccag aaacgctcgt gaaagtaaaa gacgcagagg accaattggg ggcacgagtg
     2101 ggatacatag aactggactt gaatagcggt aaaatccttg agagttttcg ccctgaagag
     2161 cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gagcagtatt atcccgtgta
     2221 gatgcggggc aagagcaact cggacgacga atacactatt cgcagaatga cttggttgaa
     2281 tactccccag tgacagaaaa gcaccttacg gacggaatga cggtaagaga attatgtagt
     2341 gccgccataa cgatgagtga taacactgcg gcgaacttac ttctgacaac catcggtgga
     2401 ccgaaggaat taaccgcttt tttgcacaat atgggagacc atgtaactcg ccttgaccgt
     2461 tgggaaccag aactgaatga agccatacca aacgacgagc gagacaccac aatgcctgcg
     2521 gcaatggcaa caacattacg caaactatta actggcgaac tacttactct ggcttcacgg
     2581 caacaattaa tagactggct tgaagcggat aaagttgcag gaccactact gcgttcggca
     2641 cttcctgctg gctggtttat tgctgataaa tctggggcag gagagcgtgg ttcacggggt
     2701 atcattgccg cacttggacc agatggtaag ccttcccgta tcgtagttat ctacacgacg
     2761 ggtagtcagg caactatgga cgaacgaaat agacagattg ctgaaatagg ggcttcactg
     2821 attaagcatt ggtaaaccga tacaattaaa ggctcctttt ggagcctttt tttttggacg
     2881 gaccggtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2941 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3001 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3061 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3121 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3181 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3241 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3301 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3361 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3421 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3481 ggcggagcct atggaaa
//
