LOCUS       sequence-330483-        4157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Containing His-ELP-intein multifunctional tag for protein 
            purification by combining Ni-NTA-based and ELP-based strategy with a
            specially designed purification protocol..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4157)
  TITLE     Yingfeng An Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4157)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pANY8
FEATURES             Location/Qualifiers
     source          1..4157
                     /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             87..104
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             126..146
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2003..2308
                     /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     2187..2206
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(2386..2404)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2400..2447
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2515..2520,2521..2537,2538..2543))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(2556..3371)
                     /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(2556..3371)
                     /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     3279..3298
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3363..3382
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3493..4081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3982..4001
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgggccacc atcatcacca ccatgattat gatattccaa
      121 ctaccgagaa tttgtatttt cagggggtcc cggggaaagg cgtgcctggt gtcggggttc
      181 caggtgtggg tgtaccaggg gtaggtgtac caggtgtggg cgttccaggc gttggtgtac
      241 caggagtagg agtcccaggt gtaggcgtac cgggttttgg agtacctgga aagggcgtcc
      301 caggggttgg tgtacccggc gttggtgttc ctggggtagg tgttccaggc gtaggagtac
      361 caggcgttgg agttcctggt gttggggtgc ctggcgttgg ggttcctggt tttggtgtcc
      421 caggaaaggg agttcctggc gtaggcgttc caggagtcgg cgtaccggga gttggagttc
      481 caggagtagg cgttcctgga gtcggagtac ctggggtcgg cgtgcctgga gtcggtgttc
      541 caggattcgg tgttcctggt aaaggcgtcc caggagtggg tgttcctggc gtcggagtgc
      601 ctggagtagg ggttcccggt gtaggcgtgc ccggagtagg tgtacctgga gttggtgtac
      661 cgggagtagg tgtccctggt ttcggagtcc ctggcaaggg ggtacccgga gttggcgtac
      721 cgggggtcgg ggtcccaggc gtcggtgtac cgggcgttgg agtccccggt gtcggtgtgc
      781 ccggcgtcgg cgtgcccggt gtaggtgtgc ctggtttcgg tgtgccaggc aagggagtac
      841 caggagtcgg ggtaccgggt gttggcgtac ccggggtggg ggttcctgga gtgggagttc
      901 cgggggtggg agtgcctggg gtgggtgtac cgggggttgg ggtgccagga ttcggagtgc
      961 cggggaaagg tgttccgggt gtgggagtcc ccggcgtagg ggtacctggt gtaggggtac
     1021 caggcgtagg tgtacccggg gtcggagttc caggtgtagg tgtaccgggt gtcggggtcc
     1081 ccggattcgg tgtgagctcg aacaacaaca acaataacaa taacaacaac gaattcgccc
     1141 tcgcagaggg cactcggatc ttcgatccgg tcaccggtac aacgcatcgc atcgaggatg
     1201 ttgtcggtgg gcgcaagcct attcatgtcg tggctgctgc caaggacgga acgctgcgtg
     1261 cgcggcccgt ggtgtcctgg ttcgaccagg gaacgcggga tgtgatcggg ttgcggatcg
     1321 ccggtggcgc catcctgtgg gcgacacccg atcacaaggt gctgacagag tacggctggc
     1381 gtgccgccgg ggaactccgc aagggagaca gggtggcgca accgcgacgc ttcgatggat
     1441 tcggtgacag tgcgccgatt ccggcgcgcg tgcaggcgct cgcggatgcc ctggatgaca
     1501 aattcctgca cgacatgctg gcggaagaac tccgctattc cgtgatccga gaagtgctgc
     1561 caacgcggcg ggcacggacg ttcggcctcg aggtcgagga actgcacacc ctcgtcgccg
     1621 aaggggttgt tgtacacaac ggatccacta gtaggcctgt cgacggtcag tccggcttac
     1681 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata
     1741 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt
     1801 gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt
     1861 ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg
     1921 gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct
     1981 gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg
     2041 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt
     2101 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     2161 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     2221 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     2281 cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtcgac
     2341 tgaccgtcag gcctccatgg gagctcctgc agttggctgc tgccaccgct gagcaataac
     2401 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     2461 ctatatccgg atatccctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa
     2521 gcttatcgat gataagctgt caaacatgag aattcttaga aaaactcatc gagcatcaaa
     2581 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2641 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2701 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2761 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     2821 ttgtgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2881 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2941 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     3001 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     3061 ttcccgggga tcgcagtggt gagtaaccat gcgtcatcag gagtacggat aaaatgcttg
     3121 atggtcggaa gcggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     3181 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     3241 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3301 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     3361 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     3421 gacaaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3481 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3541 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3601 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3661 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3721 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3781 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3841 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3901 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3961 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4021 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4081 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt tggtaccaga
     4141 tctcgatccc gcgaaat
//
