LOCUS       sequence-218117-        2662 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 2662)
  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 2662)
  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..2662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             447..752
                     /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     631..650
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             828..845
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(892..910)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      906..953
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1021..1026,1027..1043,1044..1049))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(1062..1877)
                     /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(1062..1877)
                     /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     1785..1804
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1869..1888
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1998..2586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2487..2506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatacgact cactataggg agacgacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgactagta ggcctgtcga ccctgggtct ggagaccggc
      121 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
      181 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
      241 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
      301 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
      361 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
      421 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
      481 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
      541 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
      601 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
      661 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
      721 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
      781 cggtctctcg tcgacctatg gtcaggcctg gatccgagct cctgcagcac caccaccacc
      841 accactaagg ctgctaacaa agcccgaaag aagctgagtt ggctgctgcc accgctgagc
      901 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
      961 gaggaactat atccggatat ccctgactgc gttagcaatt taactgtgat aaactaccgc
     1021 attaaagctt atcgatgata agctgtcaaa catgagaatt cttagaaaaa ctcatcgagc
     1081 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     1141 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     1201 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     1261 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     1321 aaaagtttgt gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     1381 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     1441 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     1501 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     1561 gctgttttcc cggggatcgc agtggtgagt aaccatgcgt catcaggagt acggataaaa
     1621 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     1681 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     1741 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     1801 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc
     1861 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     1921 gttcatgacc aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1981 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2041 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2101 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2161 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2221 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2281 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2341 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2401 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2461 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2521 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2581 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gcctttggta
     2641 ccagatctcg atcccgcgaa at
//
