LOCUS       sequence-218115-        3732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid is used for efficient cloning of DNA and 
            temperature-induced expression of protein in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3732)
  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 3732)
  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: pANY3
FEATURES             Location/Qualifiers
     source          1..3732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             370..675
                     /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     554..573
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             751..768
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(815..833)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      829..876
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(944..949,950..966,967..972))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(985..1800)
                     /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(985..1800)
                     /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     1708..1727
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1792..1811
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1921..2509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2410..2429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2590..3303)
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             3392..3400
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    3649..3663
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
ORIGIN
        1 catatgacta gtaggcctgt cgaccctggg tctggagacc ggcttactaa aagccagata
       61 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
      121 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
      181 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
      241 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
      301 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
      361 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
      421 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
      481 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
      541 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
      601 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
      661 ttctggggaa tataaatgtc aggctccctt atacacagcc agtcggtctc tcgtcgacct
      721 atggtcaggc ctggatccga gctcctgcag caccaccacc accaccacta aggctgctaa
      781 caaagcccga aagaagctga gttggctgct gccaccgctg agcaataact agcataaccc
      841 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
      901 tatccctgac tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg
      961 ataagctgtc aaacatgaga attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     1021 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     1081 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     1141 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     1201 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tgtgcatttc
     1261 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     1321 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     1381 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     1441 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     1501 cgcagtggtg agtaaccatg cgtcatcagg agtacggata aaatgcttga tggtcggaag
     1561 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     1621 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     1681 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     1741 atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat
     1801 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg accaaatccc
     1861 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1921 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1981 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2041 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2101 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2161 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2221 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2281 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2341 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2401 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2461 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2521 cgcggccttt ttacggttcc tggccttttg ctggcctttg gtaccgcgcc gaccagaaca
     2581 ccttgccgat cagccaaacg tctcttcagg ccactgacta gcgataactt tccccacaac
     2641 ggaacaactc tcattgcatg ggatcattgg gtactgtggg tttagtggtt gtaaaaacac
     2701 ctgaccgcta tccctgatca gtttcttgaa ggtaaactca tcacccccaa gtctggctat
     2761 gcagaaatca cctggctcaa cagcctgctc agggtcaacg agaattaaca ttccgtcagg
     2821 aaagcttggc ttggagcctg ttggtgcggt catggaatta ccttcaacct caagccagaa
     2881 tgcagaatca ctggcttttt tggttgtgct tacccatctc tccgcatcac ctttggtaaa
     2941 ggttctaagc ttaggtgaga acatccctgc ctgaacatga gaaaaaacag ggtactcata
     3001 ctcacttcta agtgacggct gcatactaac cgcttcatac atctcgtaga tttctctggc
     3061 gattgaaggg ctaaattctt caacgctaac tttgagaatt tttgtaagca atgcggcgtt
     3121 ataagcattt aatgcattga tgccattaaa taaagcacca acgcctgact gccccatccc
     3181 catcttgtct gcgacagatt cctgggataa gccaagttca tttttctttt tttcataaat
     3241 tgctttaagg cgacgtgcgt cctcaagctg ctcttgtgtt aatggtttct tttttgtgct
     3301 catacgttaa atctatcacc gcaagggata aatatctaac accgtgcgtg ttgactattt
     3361 tacctctggc ggtgataatg gttgcatgta ctaaggaggt tgtatggaac aacgcataac
     3421 cctgaaagat tatgcaatgc gctttgggca aaccaagaca gctaaagatc tctcacctac
     3481 caaacaatgc ccccctgcaa aaaataaatt catataaaaa acatacagat aaccatctgc
     3541 ggtgataaat tatctctggc ggtgttgaca taaataccac tggcggtgat actgagcaca
     3601 tcagcaggac gcactgacca ccatgaaggt gacgctctta aaaattaagc cctgaagaag
     3661 ggcagcattc aaagcagaag gctttggggt gtgtgatacg aaacgaagca ttggttaaaa
     3721 attaaggagg aa
//
