LOCUS       sequence-213051-        2663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Universal mini-vector 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 2663)
  AUTHORS   Liu X, Li T, Hart DJ, Gao S, Wang H, Gao H, Xu S, Zhang Y, Liu Y, An
            Y
  TITLE     A universal mini-vector and an annealing of PCR products (APP)-based
            cloning strategy for convenient molecular biological manipulations.
  JOURNAL   Biochem Biophys Res Commun. 2018 Mar 18;497(4):978-982. doi: 
            10.1016/j.bbrc.2018.02.099. Epub 2018 Feb 12.
  PUBMED    29448102
REFERENCE   2  (bases 1 to 2663)
  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: 
            "10.1016/j.bbrc.2018.02.099"; journalName: "Biochem Biophys Res 
            Commun"; date: "2018-03-18- 18"; volume: "497"; issue: "4"; pages: 
            "978-982"
COMMENT     Sequence Label: pANY1
FEATURES             Location/Qualifiers
     source          1..2663
                     /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             509..814
                     /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     693..712
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     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      1999..2587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2488..2507
                     /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 caggggggat ccactagtag gcctgtcgac ggtcagtccg
      181 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
      241 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
      301 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
      361 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
      421 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
      481 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
      541 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
      601 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
      661 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
      721 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
      781 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
      841 gtcgactgac cgtcaggcct ccatgggagc tcctgcagtt 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 tcggaagcgg 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 gttcatgaca aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1981 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2041 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2101 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2161 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2221 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2281 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2341 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2401 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2461 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2521 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2581 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggcctttggt
     2641 accagatctc gatcccgcga aat
//
