LOCUS       sequence-213564-        4094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid is used for efficient cloning of DNA, efficient 
            expression of protein in E. coli, and ELP-based protein 
            purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4094)
  AUTHORS   Gao H, Qi X, Hart DJ, Gao S, Wang H, Xu S, Zhang Y, Liu X, Liu Y, An
            Y
  TITLE     Three Novel Escherichia coli Vectors for Convenient and Efficient 
            Molecular Biological Manipulations.
  JOURNAL   J Agric Food Chem. 2018 Jun 20;66(24):6123-6131. doi: 
            10.1021/acs.jafc.8b01960. Epub 2018 Jun 11.
  PUBMED    29798665
REFERENCE   2  (bases 1 to 4094)
  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: 
            "10.1021/acs.jafc.8b01960"; journalName: "J Agric Food Chem"; date: 
            "2018-06-20- 20"; volume: "66"; issue: "24"; pages: "6123-6131"
COMMENT     Sequence Label: pANY6
FEATURES             Location/Qualifiers
     source          1..4094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      53..641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     542..561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     718..737
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        718..736
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             768..790
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2657..2962
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2841..2860
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(3040..3058)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3054..3101
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3169..3174,3175..3191,3192..3197))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(3210..4025)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPVPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3933..3952
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4017..4036
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
       61 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      121 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      181 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
      241 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      301 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      361 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      421 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      481 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      541 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      601 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      661 ttttacggtt cctggccttt tgctggcctt tggtaccaga tctcgatccc gcgaaattaa
      721 tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt
      781 aagaaggaga tatacatatg gtcccgggga aaggcgtgcc tggtgtcggg gttccaggtg
      841 tgggtgtacc aggggtaggt gtaccaggtg tgggcgttcc aggcgttggt gtaccaggag
      901 taggagtccc aggtgtaggc gtaccgggtt ttggagtacc tggaaagggc gtcccagggg
      961 ttggtgtacc cggcgttggt gttcctgggg taggtgttcc aggcgtagga gtaccaggcg
     1021 ttggagttcc tggtgttggg gtgcctggcg ttggggttcc tggttttggt gtcccaggaa
     1081 agggagttcc tggcgtaggc gttccaggag tcggcgtacc gggagttgga gttccaggag
     1141 taggcgttcc tggagtcgga gtacctgggg tcggcgtgcc tggagtcggt gttccaggat
     1201 tcggtgttcc tggtaaaggc gtcccaggag tgggtgttcc tggcgtcgga gtgcctggag
     1261 taggggttcc cggtgtaggc gtgcccggag taggtgtacc tggagttggt gtaccgggag
     1321 taggtgtccc tggtttcgga gtccctggca agggggtacc cggagttggc gtaccggggg
     1381 tcggggtccc aggcgtcggt gtaccgggcg ttggagtccc cggtgtcggt gtgcccggcg
     1441 tcggcgtgcc cggtgtaggt gtgcctggtt tcggtgtgcc aggcaaggga gtaccaggag
     1501 tcggggtacc gggtgttggc gtacccgggg tgggggttcc tggagtggga gttccggggg
     1561 tgggagtgcc tggggtgggt gtaccggggg ttggggtgcc aggattcgga gtgccgggga
     1621 aaggtgttcc gggtgtggga gtccccggcg taggggtacc tggtgtaggg gtaccaggcg
     1681 taggtgtacc cggggtcgga gttccaggtg taggtgtacc gggtgtcggg gtccccggat
     1741 tcggtgtgag ctcgaacaac aacaacaata acaataacaa caacgaattc gccctcgcag
     1801 agggcactcg gatcttcgat ccggtcaccg gtacaacgca tcgcatcgag gatgttgtcg
     1861 gtgggcgcaa gcctattcat gtcgtggctg ctgccaagga cggaacgctg cgtgcgcggc
     1921 ccgtggtgtc ctggttcgac cagggaacgc gggatgtgat cgggttgcgg atcgccggtg
     1981 gcgccatcct gtgggcgaca cccgatcaca aggtgctgac agagtacggc tggcgtgccg
     2041 ccggggaact ccgcaaggga gacagggtgg cgcaaccgcg acgcttcgat ggattcggtg
     2101 acagtgcgcc gattccggcg cgcgtgcagg cgctcgcgga tgccctggat gacaaattcc
     2161 tgcacgacat gctggcggaa gaactccgct attccgtgat ccgagaagtg ctgccaacgc
     2221 ggcgggcacg gacgttcggc ctcgaggtcg aggaactgca caccctcgtc gccgaagggg
     2281 ttgttgtaca caacggatcc actagtaggc ctgtcgacgg tcagtccggc ttactaaaag
     2341 ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata
     2401 tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt
     2461 tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta
     2521 agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat
     2581 caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag
     2641 aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     2701 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc
     2761 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     2821 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     2881 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     2941 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt cgactgaccg
     3001 tcaggcctcc atgggagctc ctgcagttgg ctgctgccac cgctgagcaa taactagcat
     3061 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     3121 ccggatatcc ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat
     3181 cgatgataag ctgtcaaaca tgagaattct tagaaaaact catcgagcat caaatgaaac
     3241 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     3301 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     3361 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     3421 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttgtgc
     3481 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     3541 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     3601 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     3661 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     3721 gggatcgcag tggtgagtaa ccatgcgtca tcaggagtac ggataaaatg cttgatggtc
     3781 ggaacaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     3841 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     3901 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     3961 tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg
     4021 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
     4081 aatcccttaa cgtg
//
