LOCUS       sequence-213560-        2664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid is used for efficient cloning of DNA and expression of 
            protein in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2664)
  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 2664)
  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: pANY2
FEATURES             Location/Qualifiers
     source          1..2664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(66..84)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      80..127
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(195..200,201..217,218..223))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(236..1051)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(236..1051)
                     /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
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     959..978
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1043..1062
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1173..1761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1662..1681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1838..1857
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1838..1856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1888..1910
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2284..2589
                     /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     2468..2487
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 caccaccacc accaccacta aggctgctaa caaagcccga aaggaagctg agttggctgc
       61 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
      121 ttttttgctg aaaggaggaa ctatatccgg atatccctga ctgcgttagc aatttaactg
      181 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttaga
      241 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
      301 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
      361 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
      421 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
      481 ccggtgagaa tggcaaaagt ttgtgcattt ctttccagac ttgttcaaca ggccagccat
      541 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
      601 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
      661 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
      721 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcgtcatcag
      781 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
      841 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
      901 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
      961 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     1021 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     1081 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1141 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1201 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1261 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     1321 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1381 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1441 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1501 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1561 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1621 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1681 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     1741 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     1801 tgctggcctt tggtaccaga tctcgatccc gcgaaattaa tacgactcac tatagggaga
     1861 cgacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     1921 actagtaggc ctgtcgaccc tgggtctgga gaccggctta ctaaaagcca gataacagta
     1981 tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag
     2041 tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc
     2101 tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc
     2161 agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg
     2221 ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt
     2281 gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
     2341 gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca
     2401 cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa
     2461 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa
     2521 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
     2581 ggaatataaa tgtcaggctc ccttatacac agccagtcgg tctctcgtcg acctatggtc
     2641 aggcctggat ccgagctcct gcag
//
