LOCUS       sequence-12813-e        8379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8379)
  AUTHORS   Kim JH, Wei JR, Wallach JB, Robbins RS, Rubin EJ, Schnappinger D
  TITLE     Protein inactivation in mycobacteria by controlled proteolysis and 
            its application to deplete the beta subunit of RNA polymerase.
  JOURNAL   Nucleic Acids Res. 2011 Mar;39(6):2210-20. doi: 10.1093/nar/gkq1149
  PUBMED    21075796
REFERENCE   2  (bases 1 to 8379)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2011-03"; volume: "39"; issue: "6"; 
            pages: "2210-20"
COMMENT     Sequence Label: pGMCZ-PrpoB
FEATURES             Location/Qualifiers
     source          1..8379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..104
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     101..117
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     101..117
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    161..185
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4780..4804)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4810..4830)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        4893..4911
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4893..4910
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5097..5116)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5098..5116)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5199..5787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5279..5298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5864..5950)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5992..6366)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(6385..6432)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     6464..6483
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 catgaggtna ccatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
       61 gcctttgagt gagctgatac cagcggataa caatttcaca caggaaacag ctatgaccat
      121 gattacgcca agctatcaac tttgtataga aaagttgaat tcaagtttgt acaaaaaagc
      181 aggctcggta tgtcggagga gaaggacgag gccaccgcgg ccgccgagca ggccctggtg
      241 gacgccggcc agcacgacgg cggtgtcgag gagatcgagg gtgtgccgcc gcagctgcag
      301 gccaccccgg gtatgcccga gcgcaaggcc gtggcccgcc gcaaggcccg cgtgcgcgag
      361 atcctgcaca ccctcccgcc cgccgcgcag gccgcgatcc tcgaagagct cgaccgcgac
      421 cagccgcagc tgtccgcccc gaccacccag acggccgcca ccgcgcccgt cgagaactac
      481 gacgactcgc cgacgggcgt catcgaggtg cccaagcagg cgtgagtttg ccgcgatcag
      541 gccgggccgt cacggcccgg cctgacgtgt ctcaaggggc cggcgcgcgg gcaaccgcgt
      601 cggccccacc cgccgcgatt tcggcgattt tctccggatt tccacgcccg caacgatttt
      661 cgctaagatg acggaggtgc gtgcagccgg ccggtcgggt cgtcgcacgc accggcgggt
      721 gccggtccac acgacggtgc cggccggaac gggaccgtac gacgaccgcg ggcacgcggg
      781 agtgggcgga cgcacccaag atcggcggtt gccggtcgtg gcccgggtgt gccggcgcgc
      841 tgggtggcgg gtggaacgtc acgcgagcgc aattcccccc accaaacagc gtgttcgctc
      901 ttgacggacg gccccggacg ggccaatctg ttgggcagca tgttcgaacg ggttgaggac
      961 gccagccagc gccgggtgac ccagctcatg cgcccgaggg gtttggttga ggaatgcgcc
     1021 gttctgcgct atcgttggac gttgcgctgg ctgcaccctg cccacctcac ctgtacctga
     1081 caccgtggtc ctagcctgag tagtttgctc agtacctggt cctgtgtcgt tgcgtccagg
     1141 gttctggaca ggcccaagcc agccgaaccg acgcagaatc gcgacagaga tccggcgaat
     1201 gccgcatccg aatagtcgca tgaggtgctg gaaggatgca tcttggcagt ctctagccag
     1261 agcaagtcga acgctatcac caataactcc gtcccagggg caccgaaccg agtttcattt
     1321 gccaagctcc gtgaaccgct tgaggttccg gggctgctcg acgttcagac ggattccttc
     1381 gagtggctgg tcgggtccga ccggtggcgg caggccgcca tcgaccgcgg cgaggagaac
     1441 cccgtcggcg gcctcgaaga ggtcctcgcc gagctttccc ccatcgagga tttctcgggc
     1501 tcgatgtcac tgtccttctc ggatccgcgc ttcgacgagg tcaaggcctc ggtcgacgag
     1561 tgcaaagaca aggacatgac gtacgcggcc ccgctgttcg tcacggctga gttcatcaac
     1621 aacaacaccg gcgagatcaa gagccagacg gtcttcatgg gtgacttccc gatgatgacc
     1681 gaaaagggca ccttcatcat caacggcacc gagcgcgtcg tggtgtcgca gctggtccgt
     1741 tcgcctggcg tgtacttcga cgagaccatc gacaagtcca ccgagaagac gctgcacagc
     1801 gtcaaggtga tcccgggccg cggtgcgtgg ctggagttcg acgtcgacaa gcgcgacacc
     1861 gtcggtgtgc gtatcgaccg caagcgccgc cagccggtca ccgtgctgct caaggcgctc
     1921 ggttggacca acgagcagat cgtggagcgc ttcggcttct ccgagatcat gatgggcacg
     1981 ctggagaagg acaccacctc cggcaccgac gaggccctgc tcgacatcta ccgcaagctg
     2041 cgcccgggcg agccgccgac caaggagtcc gcgcagaccc tgctggagaa cctgttcttc
     2101 aaggagaagc gctacgacct ggcccgtgtg ggccgttaca aggtcaacaa gaagctgggc
     2161 ctgaacgcgg gcaagccgat caccagctcg acgctgaccg aagaggacgt cgtcgcgacc
     2221 atcgagtacc tggtgcgtct gcacgagggt cagacttcga tgaccgtccc gggtggcgtc
     2281 gaggttcccg tcgaggtcga cgacatcgac cacttcggca accgtcgtct gcgcaccgtc
     2341 ggtgagctga tccagaacca gatccgcgtg ggcctgtccc gcatggagcg tgtcgtgcgt
     2401 gagcgcatga ccacccagga cgtcgaggcg atcacgccgc agaccctgat caacatccgt
     2461 cccgtcgtgg cggcgatcaa ggagttcttc ggcaccagcc agctgtcgca gttcatggac
     2521 cagaacaacc cgctgtcggg tctgacccac aagcgtcgtc tttcggcgct gggccccggc
     2581 ggtctgtccc gtgagcgcgc cggcctcgag gtccgcgacg tgcaccccag ccactacggc
     2641 cgcatgtgcc cgatcgagac ccctgagggt cccaacatcg gtctgatcgg ttcgctgtcg
     2701 gtgtacgccc gcgtgaaccc gttcggcttc atcgagacgc cgtaccgcaa ggtcgagaac
     2761 ggtgtggtca ccgaccagat cgactacctg accgccgacg aggaggaccg ccacgtcgtg
     2821 gcgcaggcca actcgccgac cgacgagaac ggccgcttca ccgaggaccg cgtcatggtc
     2881 cgcaagaagg gcggcgaggt cgagttcgtc tccgccgacc aggtggacta catggacgtc
     2941 tcgccgcgcc agatggtgtc ggtcgccacg gccatgatcc cgttcctcga gcacgacgac
     3001 gccaaccgcg ccctgatggg tgccaacatg cagcgccagg cggttccgct ggtgcgcagc
     3061 gaggcaccgc tggtgggtac cggtatggaa ctgcgcgccg cgatcgacgc cggtgacgtg
     3121 gtcgtcgccg acaagaccgg tgtcatcgag gaggtctcgg ccgactacat cacggtcatg
     3181 gccgacgacg gcacgcgtca gtcctaccgc ctgcgcaagt tcgcccggtc caaccacggc
     3241 acgtgcgcca accagcgtcc gatcgtggac gccgggcagc gtgtcgaggc cggccaggtc
     3301 atcgccgacg gcccctgcac ccagaacggt gagatggccc tgggcaagaa cctgctcgtg
     3361 gcgatcatgc cgtgggaagg ccacaactac gaggacgcga tcatcctctc gaaccgcctg
     3421 gtcgaagagg acgtgctcac ctcgatccac atcgaggagc acgagatcga tgcccgcgac
     3481 accaagctgg gcgccgagga gatcacccgg gacatcccga acgtctccga cgaggtgctc
     3541 gctgacctcg acgagcgcgg catcgtccgc atcggcgccg aggtccgcga cggtgacatc
     3601 ctggtcggca aggtcacccc gaagggcgag accgagctga ccccggagga gcggctgctc
     3661 cgcgcgatct tcggtgagaa ggcgcgcgag gtccgcgaca cgtcgctgaa ggtcccgcac
     3721 ggtgagtcgg gcaaggtcat cggcatccgc gtgttcagcc gcgaggacga cgacgagctg
     3781 cccgccggtg tcaacgagct ggtccgcgtc tacgtcgccc agaagcgcaa gatctccgac
     3841 ggcgacaagc tcgccggacg ccacggcaac aagggcgtca tcggcaagat cctgcccgtc
     3901 gaggacatgc cgttcctgcc ggacggcacg ccggtcgaca tcatcctgaa cacccacggt
     3961 gtgccgcgtc gtatgaacat cggccagatc ctggagaccc acctcgggtg ggtggccaag
     4021 gccggctgga acatcgacgt cgccgcgggc gtgccggact gggcgtccaa gctgcccgag
     4081 gagctgtact cggctcccgc ggacagcacg gtggcgaccc cggtgttcga cggcgcgcag
     4141 gaaggcgagc tcgccggtct gctcggctcg accctgccga accgcgacgg tgaggtcatg
     4201 gtcgacgccg acggcaagtc gacgctgttc gacggccgca gtggtgaacc gttcccgtac
     4261 ccggtgacgg ttggctacat gtacatcctg aagctgcacc acctggtcga cgacaagatc
     4321 cacgcgcgtt cgaccggtcc gtactcgatg atcacccagc agccgctcgg tggtaaggcg
     4381 cagttcggtg gtcagcgctt cggcgagatg gaatgttggg ccatgcaggc gtacggcgct
     4441 gcctacacgc tgcaggaact gctgaccatc aagtccgacg acacggtggg ccgcgtcaag
     4501 gtgtacgagg cgatcgtcaa gggcgagaac atccctgaac ccggtatccc cgagtcgttc
     4561 aaggtgttgc tcaaggagct gcagtcgctg tgcctcaacg tcgaggtgct ctccagcgac
     4621 ggcgcggcga tcgagatgcg tgacggtgac gacgaggacc tggagcgcgc tgccgcgaac
     4681 ctgggaatca acctgtcgcg caacgaatcc gcgtccgtcg aggatctcgc gtagagcctc
     4741 gtccgaagac cgcctcagta gtcaagtttc aacacccgca cccagctttc ttgtacaaag
     4801 tggacgcgtc aactttatta tacatagttg cttaagctgc aggggaccct agaggtcccc
     4861 ttttttattt taaaaatttt ttataaccgg tgatttaggt gacactatag aagtggaatt
     4921 cgatggccta gctggcatcg gtacgtgccg cgcaaccatg tagtagtcct ggagcgtgtc
     4981 catctggtgt tcaagctttc ggctagatgt gcaacgcatg gcagtcgcca atccacggcc
     5041 gaagcagagc gctatttcac tgacgcgtgc atgccatatg gaaaccgttg tggtctccct
     5101 atagtgagtc gtattaattt acgcgtttca ctggccgaca tgtgagcaaa aggccagcaa
     5161 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5221 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5281 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5341 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5401 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5461 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5521 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5581 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5641 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5701 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5761 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5821 gctcagtgga actagtcgca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     5881 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     5941 gttttatttg atgcctggca gttccctact ctcgcatggg gagagatgca ttcagtcctg
     6001 ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc
     6061 ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga
     6121 cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag
     6181 ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg
     6241 gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca
     6301 gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt
     6361 ggccatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
     6421 ttaattgtca acacgtgctc gagcaagacg tttcccgttg aatatggctc ataacacccc
     6481 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     6541 gtgcaatgta acatcagaga ttttgagaca caacgtcgct ttgttgctag ctggcgggaa
     6601 cggcaatgct cctggcggcg gcggggctgg tgcccaggtc tccgcacaga gcggcggtgc
     6661 tggcgctcgc ggccaggcgt ggttcttcgc gtactgacaa gaaacccccc tctttaggac
     6721 tcagtgtcct tgggaggggg gctttttgcg tttcaggagg tcttggccag cttggacatc
     6781 gcctcagcga tagcctcgtc gcgggcctca gacgccatct ggtacttcat cgccatccta
     6841 ggagtcgtgt gaccgagacg ggccatcagc tccttggtcg tcgcacctgc ctgagcggcg
     6901 aacgtagcgc cgacagcgcg gaggtcgtgg atgcggagtt ccggccgacc gatcttggcg
     6961 tagccacgct tcagcgactt ggtgaacgcg gacttcgaca gccggttgcc ctgcgtcgtg
     7021 gtcaccagga atgcctcggg gcccttgttc atcttcgtac ggtccttcat gtgcgctcgg
     7081 atcatctccg cgacgtgagg cggaaccgtc acaggacgct tcgaccggac ggtcttggcg
     7141 ttgccaacga cgatcttgtt ccccacgcgg gaagcgccac ggcgcacccg gagcttcatc
     7201 gtcatgccgt cgtccacgat gtccttgcgg cgaagctcga tcagctctcc gaaccggagg
     7261 ctcgtccacg ccaggatgta tgccgcgatc cggtagtgct cgaagatctc agcggcgacg
     7321 atgtccagct cctcaggcgt cagcgcctct acgtcgcgct catcggctgc cttctgctcg
     7381 atccggcacg ggttctctgc gatcagcttg tcctcgaccg ctgtgttcat caccgcccgg
     7441 aggacgttgt aggcatgccg gcgggcagtc gggtgcttcc tacccatccc ggcccaccac
     7501 gcacgcacca gagctggcgt catctctgtg accgccactt cacctagcac cgggtagatg
     7561 cggcgctccg cgtgcccgct gtacagatcc ctggtgccgt ctgcgaggtc gcgctccacg
     7621 agccacttcc gggtgtactc ctccagcgtg atggcgctgg cggctgcctt cttcgcccgg
     7681 tcctgtggag gggtccaggt ctccatctcg atgagccgct tctcgcccgc gagccaggct
     7741 tcggcgtcca tcttgttgtc gtaggtctgc agcgcgtagt acctcacacc gtcctgcggg
     7801 ttgacgtatg aggcttggat cctcccgctg cgctgagtct tcagcgatcc ccatccgcga
     7861 cgtgccaact aggtctcctc tcgtcgtgaa caaggctacc gggttgcaac tcctgtgcaa
     7921 ctctcaggct tcaacgcgct tctacgacct gcaatttctt tccacttaga ggatgcagcc
     7981 gagagggggt aaaaacctat cttgaccggc ccatatatgt ggtcggcaga cacccattct
     8041 tccaaactag ctacgcgggt tcgattcccg tcgcccgctc cgctggtcag agggtgtttt
     8101 cgccctctgg ccatttttct ttccaggggt ctgcaactct tgtgcgactc ttctgacctg
     8161 ggcatacgcg gttgcaacgc atccctgatc tggctacttt cgatgctgac aaacgaatag
     8221 agccccccgc ctgcgcgaac agacgagggg cattcacacc agattggagc tggtgcagtg
     8281 aagagaatag accgggacaa ggttgcaccg ggagttgcag cggtcggaac cctcgccgtc
     8341 ggcgggctgg cgttcgccct gtcgttcacg gctctagca
//
