LOCUS       sequence-218116-        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"
     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             1940..2245
                     /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     2124..2143
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(2323..2341)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2337..2384
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2452..2457,2458..2474,2475..2480))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(2493..3308)
                     /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(2493..3308)
                     /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     3216..3235
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3300..3319
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3430..4018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3919..3938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atggtcccgg ggaaaggcgt gcctggtgtc ggggttccag
      121 gtgtgggtgt accaggggta ggtgtaccag gtgtgggcgt tccaggcgtt ggtgtaccag
      181 gagtaggagt cccaggtgta ggcgtaccgg gttttggagt acctggaaag ggcgtcccag
      241 gggttggtgt acccggcgtt ggtgttcctg gggtaggtgt tccaggcgta ggagtaccag
      301 gcgttggagt tcctggtgtt ggggtgcctg gcgttggggt tcctggtttt ggtgtcccag
      361 gaaagggagt tcctggcgta ggcgttccag gagtcggcgt accgggagtt ggagttccag
      421 gagtaggcgt tcctggagtc ggagtacctg gggtcggcgt gcctggagtc ggtgttccag
      481 gattcggtgt tcctggtaaa ggcgtcccag gagtgggtgt tcctggcgtc ggagtgcctg
      541 gagtaggggt tcccggtgta ggcgtgcccg gagtaggtgt acctggagtt ggtgtaccgg
      601 gagtaggtgt ccctggtttc ggagtccctg gcaagggggt acccggagtt ggcgtaccgg
      661 gggtcggggt cccaggcgtc ggtgtaccgg gcgttggagt ccccggtgtc ggtgtgcccg
      721 gcgtcggcgt gcccggtgta ggtgtgcctg gtttcggtgt gccaggcaag ggagtaccag
      781 gagtcggggt accgggtgtt ggcgtacccg gggtgggggt tcctggagtg ggagttccgg
      841 gggtgggagt gcctggggtg ggtgtaccgg gggttggggt gccaggattc ggagtgccgg
      901 ggaaaggtgt tccgggtgtg ggagtccccg gcgtaggggt acctggtgta ggggtaccag
      961 gcgtaggtgt acccggggtc ggagttccag gtgtaggtgt accgggtgtc ggggtccccg
     1021 gattcggtgt gagctcgaac aacaacaaca ataacaataa caacaacgaa ttcgccctcg
     1081 cagagggcac tcggatcttc gatccggtca ccggtacaac gcatcgcatc gaggatgttg
     1141 tcggtgggcg caagcctatt catgtcgtgg ctgctgccaa ggacggaacg ctgcgtgcgc
     1201 ggcccgtggt gtcctggttc gaccagggaa cgcgggatgt gatcgggttg cggatcgccg
     1261 gtggcgccat cctgtgggcg acacccgatc acaaggtgct gacagagtac ggctggcgtg
     1321 ccgccgggga actccgcaag ggagacaggg tggcgcaacc gcgacgcttc gatggattcg
     1381 gtgacagtgc gccgattccg gcgcgcgtgc aggcgctcgc ggatgccctg gatgacaaat
     1441 tcctgcacga catgctggcg gaagaactcc gctattccgt gatccgagaa gtgctgccaa
     1501 cgcggcgggc acggacgttc ggcctcgagg tcgaggaact gcacaccctc gtcgccgaag
     1561 gggttgttgt acacaacgga tccactagta ggcctgtcga cggtcagtcc ggcttactaa
     1621 aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg
     1681 atatgtatac ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac
     1741 agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg
     1801 gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa
     1861 aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac
     1921 gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta
     1981 tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat
     2041 ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt
     2101 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc
     2161 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
     2221 taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc agtcgactga
     2281 ccgtcaggcc tccatgggag ctcctgcagt tggctgctgc caccgctgag caataactag
     2341 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     2401 tatccggata tccctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     2461 tatcgatgat aagctgtcaa acatgagaat tcttagaaaa actcatcgag catcaaatga
     2521 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     2581 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     2641 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     2701 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagtttg
     2761 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     2821 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     2881 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     2941 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     3001 ccggggatcg cagtggtgag taaccatgcg tcatcaggag tacggataaa atgcttgatg
     3061 gtcggaagcg gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     3121 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     3181 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     3241 aaatcagcat ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata
     3301 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac
     3361 aaaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3421 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3481 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3541 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3601 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3661 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3721 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3781 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3841 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3901 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3961 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4021 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggcctttgg taccagatct
     4081 cgatcccgcg aaat
//
